Selective histone deacetylase inhibitors for the treatment of human disease

ABSTRACT

Selective HDAC inhibitors, and pharmaceutical compositions that include the same, are described herein for the treatment of cancer, immunological diseases, inflammatory diseases, and neurological diseases.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a National Phase Application of International PatentApplication No. PCT/US2019/012770 filed Jan. 8, 2019 which claimspriority to U.S. Provisional Application No. 62/615,113 filed Jan. 9,2018, the entireties of which are incorporated herein by reference.

FIELD OF THE INVENTION

The invention relates generally to compounds that inhibit histonedeacetylase (HDAC) protein and more particularly, but not exclusively,to compounds that specifically inhibit HDAC6 and pharmaceuticalcompositions and methods of treating diseases that may beneficiallyutilize such compounds.

BACKGROUND OF THE INVENTION

A variety of diseases are known in the field to elude common treatmentmethods. For example, certain diseases and disorders that implicatehistone deacetylase (HDAC) proteins as pharmacological targets havecontinued to evade known therapeutics and treatment methodologies.Pan-HDAC inhibitors have multiple toxicities due to off-target effects.Molecules that may specifically target certain HDAC proteins may have agreater potential for treating disease with decreased toxicity.

Accordingly, a need exists in the field for compounds, compositions, andmethods for treating such elusive diseases and disorders, includingcertain cancers, inflammatory diseases, immunological diseases, andneurological diseases, by specifically inhibiting HDAC6.

SUMMARY OF THE INVENTION

The invention meets the needs in the field by providing specificinhibitors of HDAC proteins and may be used in the treatment of certaincancers, neurological disorders, and immunological disorders. Indeed,the compounds of the invention may be used in pharmaceuticalcompositions and methods of treatment in combating these and otherrelated diseases. In some embodiments, the compounds of the inventioninclude specific inhibitors of HDAC6.

In an embodiment, the invention includes a compound of formula I, II,III, IV, or V:

wherein R¹ may be H or optionally substituted alkyl or aryl;

R² may be H or optionally substituted alkyl or aryl;

Q may be a moiety selected from the group consisting of —NR³—(CH₂)_(n)—,—(CH₂)_(n)—NR³—, —NR³—C(═O)—(CH₂)_(n)—, —(CH₂)_(n)—C(═O)—NR³—,—C(═O)—NR³—(CH₂)_(n)—, and —(CH₂)_(n)—NR³—C(═O)—, wherein n is 0 or 1,and R³ may be H or optionally substituted alkyl or aryl;

each of R⁴, R⁵, R⁶, and R⁷ may be independently selected from the groupconsisting of H, halo, and optionally substituted alkyl, aryl, alkoxy,and aryloxy;

Z may be O, S, S(═O), or S(═O)₂;

A may be a moiety selected from the group consisting of

or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, orprodrug thereof.

In some embodiments, each of R⁴, R⁵, R⁶, and R⁷ is H. In someembodiments of formula I, II, or IV, each of R⁴, R⁵, R⁶, and R⁷ is H.

In some embodiments, Q may be —NR³—C(═O)—(CH₂)_(n)— or —(CH₂)_(n)—NR³—,wherein n is 1, and R³ is H. In some embodiments of formula I, II, orIV, Q may be —NR³—C(═O)—(CH₂)_(n)—, wherein n is 1, and R³ is H. In someembodiments of formula I, III, or V, Q may be —(CH₂)_(n)—NR³—, wherein nis 1, and R³ is H.

In some embodiments, R⁴ and R⁷ are optionally substituted alkoxy. Insome embodiments, R⁴ and R⁷ are methoxy. In some embodiments of formulaI, III, or V, R⁴ and R⁷ are optionally substituted alkoxy. In someembodiments of formula I, III, or V, R⁴ and R⁷ are methoxy.

In some embodiments, R⁵ or R⁶ is optionally substituted alkyl. In someembodiments, R⁵ or R⁶ is methyl. In some embodiments of formula I, III,or V, R⁵ or R⁶ is optionally substituted alkyl. In some embodiments offormula I, III, or V, R⁵ or R⁶ is methyl.

In some embodiments, Z is S(═O)₂ or O.

In some embodiments, the compound of formula I, II, and/or IV may beN¹-hydroxy-N⁸-(4-(N-(5-methyl-1,3,4-thiadiazol-2-yl)sulfamoyl)phenyl)octanediamide (i.e., SP-2-213 or Formula VI),

or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, orprodrug thereof.

In some embodiments, the compound of formula I, III, and/or V may be6-((2,6-dimethoxy-4-((2-methyl-[1,1′-biphenyl]-3-yl)methoxy)benzyl)amino)-N-hydroxyhexanamide(i.e., SP-2-223 or Formula VII),

or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, orprodrug thereof.

In some embodiments, the compound of formula I, III, and/or V may beN-hydroxy-6-((3-methyl-4-((2-methyl-[1,1′-biphenyl]-3-yl)methoxy)benzyl)amino)hexanamide(i.e., SP-2-225 or Formula VIII),

or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, orprodrug thereof.

In some embodiments, the compounds of formulas I, II, III, IV, V, VI,VII, and/or VIII are HDAC inhibitors. In some embodiments, the compoundsof formulas I, II, III, IV, V, VI, VII, and/or VIII are selective HDAC6inhibitors.

In an embodiment, the invention includes a pharmaceutical composition orformulation that may include an HDAC inhibitor, such as a selective HDACinhibitor (e.g., selective inhibitor of HDAC6), in an amount effectiveto treat a disease alleviated by inhibiting HDAC protein in a patient inneed thereof, and a physiologically compatible carrier medium.

In some embodiments, the pharmaceutical composition may include one ormore compounds of formulas I, II, III, IV, V, VI, VII, and/or VIII, or apharmaceutically acceptable salt, solvate, hydrate, cocrystal, orprodrug thereof.

In some embodiments, the pharmaceutical composition may include acompound of formula VI, or a pharmaceutically acceptable salt, solvate,hydrate, cocrystal, or prodrug thereof.

In some embodiments, the pharmaceutical composition may include acompound of formula VII, or a pharmaceutically acceptable salt, solvate,hydrate, cocrystal, or prodrug thereof.

In some embodiments, the pharmaceutical composition may include acompound of formula VIII, or a pharmaceutically acceptable salt,solvate, hydrate, cocrystal, or prodrug thereof.

In some embodiments, the pharmaceutical composition may include aselective HDAC6 inhibitor.

In an embodiment, the invention may include a method of treating adisease alleviated by inhibiting HDAC protein, or optionally selectivelyinhibiting HDAC6 protein, in a patient in need thereof, wherein thetreatment comprises administering a therapeutically effective amount ofone or more HDAC inhibitors that may include one or more compounds offormulas I, II, III, IV, V, VI, VII, and/or VIII, or a pharmaceuticallyacceptable salt, solvate, hydrate, cocrystal, or prodrug thereof.

In some embodiments, the one or more HDAC inhibitors include a selectiveHDAC6 inhibitor.

In some embodiments, the one or more HDAC inhibitors may be administeredin dosage unit form. In some embodiments, the dosage unit may include aphysiologically compatible carrier medium.

In some embodiments, the one or more HDAC inhibitors may include acompound of formula VI, or a pharmaceutically acceptable salt, solvate,hydrate, cocrystal, or prodrug thereof.

In some embodiments, the one or more HDAC inhibitors may include acompound of formula VII, or a pharmaceutically acceptable salt, solvate,hydrate, cocrystal, or prodrug thereof.

In some embodiments, the one or more HDAC inhibitors may include acompound of formula VIII, or a pharmaceutically acceptable salt,solvate, hydrate, cocrystal, or prodrug thereof.

In some embodiments, the disease alleviated by inhibiting HDAC proteinmay be cancer, an immunological disease, an inflammatory disease, or aneurological disease.

In some embodiments, the cancer alleviated by inhibiting HDAC activity,and optionally selectively inhibiting HDAC6 activity, may be selectedfrom the group consisting of acoustic neuroma, adenocarcinoma,angiosarcoma, astrocytoma, basal cell carcinoma, bile duct carcinoma,bladder carcinoma, brain cancer, breast cancer, brochogenic carcinoma,cervical cancer, chordoma, choriocarcinoma, colon cancer, colorectalcancer, craniopharygioma, cystadenocarcinoma, embryonal carcinoma,endotheliocarconima, ependymoma, epithelial carcinoma, esophagealcancer, Ewing's tumor, fibrosarcoma, gastric cancer, glioblastomamultiforme, glioma, head and neck cancer, hemangioblastoma, hepatoma,kidney cancer, leiomyosarcoma, liposarcoma, lung cancer,lymphangioendotheliosarcoma, lymphangiosarcoma, medullary carcinoma,medulloblastoma, melanoma, meningioma, mesothelioma, myxosarcoma, nasalcancer, neuroblastoma, oligodendroglioma, oral cancer, osteogenicsarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma,papillary carcinoma, pinealoma, prostate cancer, rabdomyosarcoma, rectalcancer, renal cell carcinoma, retinoblastoma, sarcoma, sebacaceous glandcarcinoma, seminoma, skin cancer, squamous cell carcinoma, stomachcancer, sweat gland carcinoma, synovioma, testicular cancer, small celllung carcinoma, throat cancer, uterine cancer, Wilm's tumor, bloodcancer, acute erythroleukemic leukemia, acute lymphoblastic B-cellleukemia, acute lymphoblastic T-cell leukemia, acute lymphoblasticleukemia, acute megakaryoblastic leukemia, acute monoblastic leukemia,acute myeloblastic leukemia, acute myelomonocytic leukemia, acutenonlymphocytic leukemia, acute promyelocytic leukemia, acuteundifferentiated leukemia, chronic lymphocytic leukemia, chronicmyelocytic leukemia, hairy cell leukemia, multiple myeloma, heavy chaindisease, Hodgkin's disease, multiple myeloma, non-Hodgkin's lymphoma,polycythemia vera, and Waldenstrom's macroglobulinemia. In someembodiments the cancer may be an HPV16 positive (+) cancer. In someembodiments, the HPV16 and HPV18 positive (+) cancer may include one ormore of cervical cancer and head and neck cancer. In some embodiments,when the disease treated by the methods of the invention is cancer, thecancer may be selected from those cancers listed herein.

In some embodiments, the immunological disease alleviated by inhibitingHDAC activity, and optionally selectively inhibiting HDAC6 activity, maybe selected from the group consisting of celiac disease, diabetesmellitus type 1 (IDDM), systemic lupus erythematosus (SLE), Sjogren'ssyndrome, Churg-Strauss Syndrome, Hashimoto's thyroiditis, Graves'disease, idiopathic thrombocytopenic purpura, rheumatoid arthritis (RA),polymyositis, ulcerative colitis, Crohn's disease, autoimmune carditis,Wegener's granulomatosis, autoimmune hemolytic anemia, polyarteritisnodosa, psoriasis, vitiligo, epidermolysis bullosa, scleroderma,alopecia areata, epidermolysis bullosa acquisita, bullous pemphigoid,pemphigus foliaceous, and pemphigus vulgaris.

In some embodiments, the neurological disease alleviated by inhibitingHDAC activity, and optionally selectively inhibiting HDAC6 activity, maybe selected from the group consisting of stroke, Huntington's disease,spinal muscular atrophy (SMA), Parkinson's disease, Alzheimer's,Multiple Sclerosis, and Amyotrophic Lateral Sclerosis (ALS).

In some embodiments, the pharmaceutical compositions may include anadditional therapeutic agent. In certain embodiments, the additionaltherapeutic agents may include one or more of a chemotherapeutic and animmunotherapeutic agent.

In some embodiments, the chemotherapeutic agent may include one or moreof bortezomib (Velcade®), lenalidomide (Revlimid®), temozolomide,5-fluorouracil (5-FU), 6-mercaptopurine (6-MP), Capecitabine (Xeloda®),Cladribine, Clofarabine, Cytarabine (Ara-C®), Floxuridine, Fludarabine,Gemcitabine (Gemzar®), Hydroxyurea, Methotrexate, Pemetrexed (Alimta®),Pentostatin, Thioguanine, daunorubicin; doxorubicin, epirubicin,idaurubicin, topotecan, irinotecan, etoposide, teniposide, mitoxantrone,Vinblastine, vincristine, vinorelbine, estramustine, paclitaxel,dexamethasone, and docetaxel.

In some embodiments, the immunotherapeutic agent may include a one ormore of a PD-1 inhibitor and a PD-L1 inhibitor, wherein the PD-1 andPD-L1 inhibitor may be selected from the group consisting of nivolumab,pembrolizumab, pidilizumab, durvalumab, atezolizumab, avelumab, or anyfragment, derivative, conjugate, variant, radioisotope-labeled complex,or biosimilar thereof.

In some embodiments, the immunotherapeutic agent may include one or moreof rituximab, trastuzumab, ibritumomab, tositumomab, cetuximab,bevacizumab, gemtuzumab, alemtuzumab, and BL22 or any fragment,derivative, conjugate, variant, radioisotope-labeled complex, orbiosimilar thereof.

In another embodiment, the method of the invention may be a second orthird line method of treatment for the patient and administration of thecompound occurs after performance of a first or second therapy on thepatient that failed to treat the disease.

Accordingly, as briefly described herein, this disclosure includescompounds, compositions, and methods of treatment that provide treatmentsolutions to answer the needs in the field.

BRIEF DESCRIPTION OF THE DRAWINGS

The foregoing summary and the following detailed description of theexemplary embodiments as described herein may be further understood whenread in conjunction with the appended drawings, in which:

FIG. 1 illustrates the in vitro inhibitory activity of SP-2-213,SP-2-223, and SP-2-225 against HDAC1, HDAC3, HDAC6 and pan-HDACproteins.

FIGS. 2A to 2C illustrate the in vitro inhibitory activity of SP-2-213(FIG. 2A), SP-2-223 (FIG. 2B), and SP-2-225 (FIG. 2C) against HDAC6protein.

DETAILED DESCRIPTION OF THE INVENTION

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one of skill in theart to which this invention belongs. All patents and publicationsreferred to herein are incorporated by reference in their entireties.

Definitions

As used herein, the terms “administer,” “administration” or“administering” refer to (1) providing, giving, dosing, and/orprescribing by either a health practitioner or his authorized agent orunder his or her direction according to the disclosure; and/or (2)putting into, taking or consuming by the mammal, according to thedisclosure.

The terms “co-administration,” “co-administering,” “administered incombination with,” “administering in combination with,” “simultaneous,”and “concurrent,” as used herein, encompass administration of two ormore active pharmaceutical ingredients to a subject so that both activepharmaceutical ingredients and/or their metabolites are present in thesubject at the same time. Co-administration includes simultaneousadministration in separate compositions, administration at differenttimes in separate compositions, or administration in a composition inwhich two or more active pharmaceutical ingredients are present.Simultaneous administration in separate compositions and administrationin a composition in which both agents are present are preferred.

The terms “active pharmaceutical ingredient” and “drug” include the HDACinhibitors, chemotherapeutic agents, and immunotherapeutic agentsdescribed herein and, more specifically, include the HDAC inhibitorsdescribed by formulas I, II, III, IV, V, VI, VII, and/or VIII. The terms“active pharmaceutical ingredient” and “drug” may also include thosecompounds described herein that bind HDAC protein and thereby modulateHDAC protein activity. In certain embodiments, the HDAC protein and HDACprotein activity may refer to HDAC6 protein and HDAC6 protein activity.

The term “isostere” refers to a group or molecule whose chemical and/orphysical properties are similar to those of another group or molecule. A“bioisostere” is a type of isostere and refers to a group or moleculewhose biological properties are similar to those of another group ormolecule. For example, for the HDAC inhibitors described herein, acarboxylic acid may be replaced by one of the following bioisosteres forcarboxylic acids, including, without limitation, alkyl esters (COOR),acylsulfonamides (CONR—SO₂R), hydroxamic acids (CONR—OH), hydroxamates(CONR—OR), tetrazoles, hydroxyisoxazoles, isoxazol-3-ones, andsulfonamides (SO₂NR), where each R may independently represent hydrogen,alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl,heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

The term “in vivo” refers to an event that takes place in a subject'sbody.

The term “in vitro” refers to an event that takes places outside of asubject's body. In vitro assays encompass cell-based assays in whichcells alive or dead are employed and may also encompass a cell-freeassay in which no intact cells are employed.

The term “effective amount” or “therapeutically effective amount” refersto that amount of a compound or combination of compounds as describedherein that is sufficient to effect the intended application including,but not limited to, disease treatment. A therapeutically effectiveamount may vary depending upon the intended application (in vitro or invivo), or the subject and disease condition being treated (e.g., theweight, age and gender of the subject), the severity of the diseasecondition, the manner of administration, etc. which can readily bedetermined by one of ordinary skill in the art. The term also applies toa dose that will induce a particular response in target cells (e.g., thereduction of platelet adhesion and/or cell migration). The specific dosewill vary depending on the particular compounds chosen, the dosingregimen to be followed, whether the compound is administered incombination with other compounds, timing of administration, the tissueto which it is administered, and the physical delivery system in whichthe compound is carried.

A “therapeutic effect” as that term is used herein, encompasses atherapeutic benefit and/or a prophylactic benefit. A prophylactic effectincludes delaying or eliminating the appearance of a disease orcondition, delaying or eliminating the onset of symptoms of a disease orcondition, slowing, halting, or reversing the progression of a diseaseor condition, or any combination thereof.

As used herein, the terms “treat,” “treatment,” and/or “treating” mayrefer to the management of a disease, disorder, or pathologicalcondition (e.g., cancer, neoplastic disorder, inflammatory disorder,immunological disorder, or neurological disorder) with the intent tocure, ameliorate, stabilize, prevent, or control the disease, disorder,pathological condition, or symptoms thereof. Regarding control of thedisease, disorder, or pathological condition more specifically,“control” may include the absence of disease progression, as assessed bythe response to the methods recited herein, where such response may becomplete (e.g., placing the disease in remission) or partial (e.g.,slowing the spread of cancerous cells and tissues and/or preventing,slowing, or halting metastasis). The terms “treat,” “treatment,” and/or“treating” may further encompass, with respect to the treatment ofcancer, the sensitization of cancerous cells and tissues (e.g.,neoplastic cells and tissues) to radiation and/or the protection ofnon-cancerous cells from the effects of radiation.

For example, a patient responding to the methods of treatment disclosedherein may exhibit the absence of disease progression (e.g., halting thegrowth and/or spread of neoplastic cells and tissues) over anotherpatient that does not receive the methods of treatment described herein.

As used herein, the terms “modulate” and “modulation” refer to a changein biological activity for a biological molecule (e.g., a protein, gene,peptide, antibody, and the like), where such change may relate to anincrease in biological activity (e.g., increased activity, agonism,activation, expression, upregulation, and/or increased expression) ordecrease in biological activity (e.g., decreased activity, antagonism,suppression, deactivation, downregulation, and/or decreased expression)for the biological molecule. For example, the compounds described hereinmay modulate (i.e., inhibit) HDAC protein. In some embodiments, thecompounds described herein may selectively modulate (i.e., selectivelyinhibit) a first HDAC protein as compared to a second HDAC protein. Incertain embodiments, the compounds described herein may selectivelymodulate (i.e., selectively inhibit) HDAC6 protein as compared to otherHDAC proteins including, but not limited to, HDAC1 and/or HDAC3. Incertain embodiments, the compounds described herein may selectivelymodulate (i.e., selectively inhibit) HDAC6 protein as compared to HDAC1protein. In certain embodiments, the compounds described herein mayselectively modulate (i.e., selectively inhibit) HDAC6 protein ascompared to HDAC3 protein.

In some embodiments, a selective HDAC6 inhibitor is a compound that isat least 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61,62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, or 75 times moreselective for HDAC6 than HDAC1 as measured by determining the IC₅₀values for the selective HDAC6 inhibitor in an in vitro HDAC enzymeinhibition assay comparing HDAC6 protein and HDAC1 protein, as describedherein. In some embodiments, a selective HDAC6 inhibitor is a compoundthat is about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59,60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, or 75 timesmore selective for HDAC6 than HDAC1 as measured by determining the IC₅₀values for the selective HDAC6 inhibitor in an in vitro HDAC enzymeinhibition assay comparing HDAC6 protein and HDAC1 protein, as describedherein.

In some embodiments, a selective HDAC6 inhibitor is a compound that isat least 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, or 55 times more selectivefor HDAC6 than HDAC3 as measured by determining the IC₅₀ values for theselective HDAC6 inhibitor in an in vitro HDAC enzyme inhibition assaycomparing HDAC6 protein and HDAC3 protein. In some embodiments, aselective HDAC6 inhibitor is a compound that is about 10, 11, 12, 13,14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49,50, 51, 52, 53, 54, or 55 times more selective for HDAC6 than HDAC3 asmeasured by determining the IC₅₀ values for the selective HDAC6inhibitor in an in vitro HDAC enzyme inhibition assay comparing HDAC6protein and HDAC3 protein.

The terms “QD,” “qd,” or “q.d.” mean quaque die, once a day, or oncedaily. The terms “BID,” “bid,” or “b.i.d.” mean bis in die, twice a day,or twice daily. The terms “TID,” “tid,” or “t.i.d.” mean ter in die,three times a day, or three times daily. The terms “QID,” “qid,” or“q.i.d.” mean quater in die, four times a day, or four times daily.

The term “pharmaceutically acceptable salt” refers to salts derived froma variety of organic and inorganic counter ions known in the art.Pharmaceutically acceptable acid addition salts can be formed withinorganic acids and organic acids. Preferred inorganic acids from whichsalts can be derived include, for example, hydrochloric acid,hydrobromic acid, sulfuric acid, nitric acid and phosphoric acid.Preferred organic acids from which salts can be derived include, forexample, acetic acid, propionic acid, glycolic acid, pyruvic acid,oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid,tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid andsalicylic acid. Pharmaceutically acceptable base addition salts can beformed with inorganic and organic bases. Inorganic bases from whichsalts can be derived include, for example, sodium, potassium, lithium,ammonium, calcium, magnesium, iron, zinc, copper, manganese andaluminum. Organic bases from which salts can be derived include, forexample, primary, secondary, and tertiary amines, substituted aminesincluding naturally occurring substituted amines, cyclic amines andbasic ion exchange resins. Specific examples include isopropylamine,trimethylamine, diethylamine, triethylamine, tripropylamine, andethanolamine. In some embodiments, the pharmaceutically acceptable baseaddition salt is chosen from ammonium, potassium, sodium, calcium, andmagnesium salts. The term “cocrystal” refers to a molecular complexderived from a number of cocrystal formers known in the art. Unlike asalt, a cocrystal typically does not involve hydrogen transfer betweenthe cocrystal and the drug, and instead involves intermolecularinteractions, such as hydrogen bonding, aromatic ring stacking, ordispersive forces, between the cocrystal former and the drug in thecrystal structure.

“Pharmaceutically acceptable carrier” or “pharmaceutically acceptableexcipient” or “physilogically compatible” carrier or carrier medium isintended to include any and all solvents, dispersion media, coatings,antibacterial and antifungal agents, isotonic and absorption delayingagents, and inert ingredients. The use of such pharmaceuticallyacceptable carriers or pharmaceutically acceptable excipients for activepharmaceutical ingredients is well known in the art. Remington: TheScience and Practice of Pharmacy, 20^(th) edition, A. R. Genaro et al.,Part 5, Pharmaceutical Manufacturing, pp. 669-1015 (Lippincott Williams& Wilkins, Baltimore, Md./Philadelphia, Pa.) (2000) discloses variouscarriers used in formulating pharmaceutical compositions and knowntechniques for the preparation thereof. Except insofar as anyconventional pharmaceutically acceptable carrier or pharmaceuticallyacceptable excipient is incompatible with the active pharmaceuticalingredient, its use in the therapeutic compositions of the invention iscontemplated. Additional active pharmaceutical ingredients, such asother drugs, can also be incorporated into the described compositionsand methods. Except insofar as any conventional pharmaceutical carriermedium is incompatible with either the compounds used as describedherein, such as by producing an undesirable biological effect orotherwise interacting in a deleterious manner with any othercomponent(s) of a formulation comprising such compounds or agents, itsuse is contemplated to be within the scope of this invention.

A “prodrug” refers to a derivative of a compound described herein, thepharmacologic action of which results from the conversion by chemical ormetabolic processes in vivo to the active compound. Prodrugs includecompounds wherein an amino acid residue, or a polypeptide chain of twoor more (e.g., two, three or four) amino acid residues is covalentlyjoined through an amide or ester bond to a free amino, hydroxyl orcarboxylic acid group of formulas I, II, III, IV, V, VI, VII, and/orVIII. The amino acid residues include but are not limited to the 20naturally occurring amino acids commonly designated by one or threeletter symbols but also include, for example, 4-hydroxyproline,hydroxylysine, desmosine, isodesmosine, 3-methylhistidine, beta-alanine,gamma-aminobutyric acid, citrulline, homocysteine, homoserine, ornithineand methionine sulfone. Additional types of prodrugs are alsoencompassed. For instance, free carboxyl groups can be derivatized asamides or alkyl esters (e.g., methyl esters and acetoxy methyl esters).Prodrug esters as employed herein includes esters and carbonates formedby reacting one or more hydroxyls of compounds of the method of theinvention with alkyl, alkoxy, or aryl substituted acylating agentsemploying procedures known to those skilled in the art to generateacetates, pivalates, methylcarbonates, benzoates and the like. Asfurther examples, free hydroxyl groups may be derivatized using groupsincluding but not limited to hemisuccinates, phosphate esters,dimethylaminoacetates, and phosphoryloxymethyloxycarbonyls, as outlinedin Advanced Drug Delivery Reviews, 1996, 19, 115. Carbamate prodrugs ofhydroxyl and amino groups are also included, as are carbonate prodrugs,sulfonate prodrugs, sulfonate esters and sulfate esters of hydroxylgroups. Free amines can also be derivatized to amides, sulfonamides orphosphonamides. All of the stated prodrug moieties may incorporategroups including but not limited to ether, amine and carboxylic acidfunctionalities. Moreover, any compound that can be converted in vivo toprovide the bioactive agent (e.g., a compound of formulas I, II, III,IV, V, VI, VII, and/or VIII) is a prodrug within the scope of theinvention. Various forms of prodrugs are well known in the art. Acomprehensive description of pro drugs and prodrug derivatives aredescribed in: (a) The Practice of Medicinal Chemistry, Camille G.Wermuth et al., (Academic Press, 1996); (b) Design of Prodrugs, editedby H. Bundgaard, (Elsevier, 1985); (c) A Textbook of Drug Design andDevelopment, P. Krogsgaard-Larson and H. Bundgaard, eds., (HarwoodAcademic Publishers, 1991). In general, prodrugs may be designed toimprove the penetration of a drug across biological membranes in orderto obtain improved drug absorption, to prolong duration of action of adrug (slow release of the parent drug from a prodrug, decreasedfirst-pass metabolism of the drug), to target the drug action (e.g.organ or tumor-targeting, lymphocyte targeting), to modify or improveaqueous solubility of a drug (e.g., i.v. preparations and eyedrops), toimprove topical drug delivery (e.g. dermal and ocular drug delivery), toimprove the chemical/enzymatic stability of a drug, or to decreaseoff-target drug effects, and more generally in order to improve thetherapeutic efficacy of the compounds utilized in the invention.

As used herein, the term “subject” or “patient” may refer to any mammal.Exemplary mammals include laboratory animals, including rodents such asmice, rats and guinea pigs; farm animals such as cows, sheep, pigs andgoats; pet animals such as dogs and cats; and primates such as monkeys,apes and humans. In certain embodiments, the “subject” or “patient” is ahuman subject or patient.

Unless otherwise stated, the chemical structures depicted herein areintended to include compounds which differ only in the presence of oneor more isotopically enriched atoms. For example, compounds where one ormore hydrogen atoms is replaced by deuterium or tritium, or wherein oneor more carbon atoms is replaced by ¹³C- or ¹⁴C-enriched carbons, arewithin the scope of this invention.

When ranges are used herein to describe, for example, physical orchemical properties such as molecular weight or chemical formulae, allcombinations and subcombinations of ranges and specific embodimentstherein are intended to be included. Use of the term “about” whenreferring to a number or a numerical range means that the number ornumerical range referred to is an approximation within experimentalvariability (or within statistical experimental error), and thus thenumber or numerical range may vary. The variation is typically from 0%to 15%, preferably from 0% to 10%, more preferably from 0% to 5% of thestated number or numerical range. The term “comprising” (and relatedterms such as “comprise” or “comprises” or “having” or “including”)includes those embodiments such as, for example, an embodiment of anycomposition of matter, method or process that “consist of” or “consistessentially of” the described features.

“Alkyl” refers to a straight or branched hydrocarbon chain radicalconsisting solely of carbon and hydrogen atoms, containing nounsaturation, having from one to ten carbon atoms (e.g., (C₁₋₁₀)alkyl orC₁₋₁₀ alkyl). Whenever it appears herein, a numerical range such as “1to 10” refers to each integer in the given range—e.g., “1 to 10 carbonatoms” means that the alkyl group may consist of 1 carbon atom, 2 carbonatoms, 3 carbon atoms, etc., up to and including 10 carbon atoms,although the definition is also intended to cover the occurrence of theterm “alkyl” where no numerical range is specifically designated.Typical alkyl groups include, but are in no way limited to, methyl,ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl isobutyl,tertiary butyl, pentyl, isopentyl, neopentyl, hexyl, septyl, octyl,nonyl and decyl. The alkyl moiety may be attached to the rest of themolecule by a single bond, such as for example, methyl (Me), ethyl (Et),n-propyl (Pr), 1-methylethyl (isopropyl), n-butyl, n-pentyl,1,1-dimethylethyl (t-butyl) and 3-methylhexyl. Unless stated otherwisespecifically in the specification, an alkyl group is optionallysubstituted by one or more of substituents which are independentlyheteroalkyl, acylsulfonamido, alkenyl, alkynyl, cycloalkyl,heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, trimethylsilanyl, —OR^(a), —SR^(a),—S(O)_(t)R^(a)— (where t is 1 or 2), —OC(O)—R^(a), —N(R^(a))₂,—C(O)R^(a), —C(O)OR^(a), —OC(O)N(R^(a))₂, —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a), —N(R^(a))C(O)N(R^(a))₂,N(R^(a))C(NR^(a))N(R^(a))₂, —N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2),—S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or2), or PO₃(R^(a))₂ where each R^(a) is independently hydrogen, alkyl,fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl,heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Alkylaryl” refers to an -(alkyl)aryl radical where aryl and alkyl areas disclosed herein and which are optionally substituted by one or moreof the substituents described as suitable substituents for aryl andalkyl respectively.

“Alkylhetaryl” refers to an -(alkyl)hetaryl radical where hetaryl andalkyl are as disclosed herein and which are optionally substituted byone or more of the substituents described as suitable substituents foraryl and alkyl respectively.

“Alkylheterocycloalkyl” refers to an -(alkyl) heterocycyl radical wherealkyl and heterocycloalkyl are as disclosed herein and which areoptionally substituted by one or more of the substituents described assuitable substituents for heterocycloalkyl and alkyl respectively.

An “alkene” moiety refers to a group consisting of at least two carbonatoms and at least one carbon-carbon double bond, and an “alkyne” moietyrefers to a group consisting of at least two carbon atoms and at leastone carbon-carbon triple bond. The alkyl moiety, whether saturated orunsaturated, may be branched, straight chain, or cyclic.

“Alkenyl” refers to a straight or branched hydrocarbon chain radicalgroup consisting solely of carbon and hydrogen atoms, containing atleast one double bond, and having from two to ten carbon atoms (i.e.,(C₂₋₁₀)alkenyl or C₂₋₁₀ alkenyl). Whenever it appears herein, anumerical range such as “2 to 10” refers to each integer in the givenrange—e.g., “2 to 10 carbon atoms” means that the alkenyl group mayconsist of 2 carbon atoms, 3 carbon atoms, etc., up to and including 10carbon atoms. The alkenyl moiety may be attached to the rest of themolecule by a single bond, such as for example, ethenyl (i.e., vinyl),prop-1-enyl (i.e., allyl), but-1-enyl, pent-1-enyl and penta-1,4-dienyl.Unless stated otherwise specifically in the specification, an alkenylgroup is optionally substituted by one or more substituents which areindependently alkyl, heteroalkyl, acylsulfonamido, alkenyl, alkynyl,cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, trimethylsilanyl, —OR^(a), —SR^(a),—S(O)_(t)R^(a)— (where t is 1 or 2), —OC(O)—R^(a), —N(R^(a))₂,—C(O)R^(a), —C(O)OR^(a), —OC(O)N(R^(a))₂, —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a), —N(R^(a))C(O)N(R^(a))₂,N(R^(a))C(NR^(a))N(R^(a))₂, —N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2),—S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or2), or PO₃(R^(a))₂, where each R^(a) is independently hydrogen, alkyl,fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl,heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Alkenyl-cycloalkyl” refers to an -(alkenyl)cycloalkyl radical wherealkenyl and cycloalkyl are as disclosed herein and which are optionallysubstituted by one or more of the substituents described as suitablesubstituents for alkenyl and cycloalkyl respectively.

“Alkynyl” refers to a straight or branched hydrocarbon chain radicalgroup consisting solely of carbon and hydrogen atoms, containing atleast one triple bond, having from two to ten carbon atoms (i.e.,(C₂₋₁₀)alkynyl or C₂₋₁₀ alkynyl). Whenever it appears herein, anumerical range such as “2 to 10” refers to each integer in the givenrange—e.g., “2 to 10 carbon atoms” means that the alkynyl group mayconsist of 2 carbon atoms, 3 carbon atoms, etc., up to and including 10carbon atoms. The alkynyl may be attached to the rest of the molecule bya single bond, for example, ethynyl, propynyl, butynyl, pentynyl andhexynyl. Unless stated otherwise specifically in the specification, analkynyl group is optionally substituted by one or more substituentswhich independently are: alkyl, heteroalkyl, alkenyl, alkynyl,cycloalkyl, heterocycloalkyl, hydroxamate, acylsulfonamido, aryl,arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano,trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, —OR^(a),—SR^(a), —S(O)_(t)R^(a)— (where t is 1 or 2), —OC(O)—R^(a), —N(R^(a))₂,—C(O)R^(a), —C(O)OR^(a), —OC(O)N(R^(a))₂, —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a), —N(R^(a))C(O)N(R^(a))₂,N(R^(a))C(NR^(a))N(R^(a))₂, —N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2),—S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or2), or PO₃(R^(a))₂, where each R^(a) is independently hydrogen, alkyl,fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl,heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Alkynyl-cycloalkyl” refers to an -(alkynyl)cycloalkyl radical wherealkynyl and cycloalkyl are as disclosed herein and which are optionallysubstituted by one or more of the substituents described as suitablesubstituents for alkynyl and cycloalkyl respectively.

“Acylsulfonamide” refers to the group —C(═O)NR^(a)—S(═O)R^(a), whereeach R^(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

“Carboxaldehyde” refers to a —(C═O)H radical.

“Carbonyl” refers to the group —C(═O)—. Carbonyl groups may besubstituted with the following exemplary substituents: alkyl,heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,hydroxamate, acylsulfonamido, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, trimethylsilanyl, —OR^(a), —SR^(a),—S(O)_(t)R^(a)— (where t is 1 or 2), —OC(O)—R^(a), —N(R^(a))₂,—C(O)R^(a), —NR^(a)—OR^(a)—, —C(O)O R^(a), —OC(O)N(R^(a))₂,—C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a),—N(R^(a))C(O)N(R^(a))₂, N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or 2), or PO₃(R^(a))₂, whereeach R^(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Carboxyl” refers to a —(C═O)OH radical.

“Cyano” refers to a —CN radical.

“Cycloalkyl” refers to a monocyclic or polycyclic radical that containsonly carbon and hydrogen, and may be saturated, or partiallyunsaturated. Cycloalkyl groups include groups having from 3 to 10 ringatoms (i.e. (C₃₋₁₀)cycloalkyl or C₃₋₁₀ cycloalkyl). Whenever it appearsherein, a numerical range such as “3 to 10” refers to each integer inthe given range—e.g., “3 to 10 carbon atoms” means that the cycloalkylgroup may consist of 3 carbon atoms, etc., up to and including 10 carbonatoms. Illustrative examples of cycloalkyl groups include, but are notlimited to the following moieties: cyclopropyl, cyclobutyl, cyclopentyl,cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl,cyclononyl, cyclodecyl, norbornyl, and the like. Unless stated otherwisespecifically in the specification, a cycloalkyl group is optionallysubstituted by one or more substituents which independently are: alkyl,heteroalkyl, alkenyl, alkynyl, cycloalkyl, acylsulfonamido,heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, trimethylsilanyl, —OR^(a), —SR^(a),—S(O)_(t)R^(a)— (where t is 1 or 2), —S(O)_(t)R^(a)— (where t is 1 or2), —OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —OC(O)N(R^(a))₂,—C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a),—N(R^(a))C(O)N(R^(a))₂, N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or 2), or PO₃(R^(a))₂, whereeach R^(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Cycloalkyl-alkenyl” refers to a -(cycloalkyl)alkenyl radical wherecycloalkyl and alkenyl are as disclosed herein and which are optionallysubstituted by one or more of the substituents described as suitablesubstituents for cycloalkyl and alkenyl, respectively.

“Cycloalkyl-heterocycloalkyl” refers to a -(cycloalkyl)heterocycloalkylradical where cycloalkyl and heterocycloalkyl are as disclosed hereinand which are optionally substituted by one or more of the substituentsdescribed as suitable substituents for cycloalkyl and heterocycloalkyl,respectively.

“Cycloalkyl-heteroaryl” refers to a -(cycloalkyl)heteroaryl radicalwhere cycloalkyl and heteroaryl are as disclosed herein and which areoptionally substituted by one or more of the substituents described assuitable substituents for cycloalkyl and heteroaryl, respectively.

The term “alkoxy” refers to the group —O-alkyl, including from 1 to 8carbon atoms of a straight, branched, cyclic configuration andcombinations thereof attached to the parent structure through an oxygen.Examples include, but are not limited to, methoxy, ethoxy, propoxy,isopropoxy, cyclopropyloxy and cyclohexyloxy. “Lower alkoxy” refers toalkoxy groups containing one to six carbons.

The term “substituted alkoxy” refers to alkoxy wherein the alkylconstituent is substituted (i.e., —O-(substituted alkyl)). Unless statedotherwise specifically in the specification, the alkyl moiety of analkoxy group is optionally substituted by one or more substituents whichindependently are: alkyl, heteroalkyl, alkenyl, acylsulfonamido,alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl,heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, trimethylsilanyl, —OR^(a), —SR^(a),—S(O)_(t)R^(a)— (where t is 1 or 2), —OC(O)—R^(a), —N(R^(a))₂,—C(O)R^(a), —C(O)OR^(a), —OC(O)N(R^(a))₂, —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a), —N(R^(a))C(O)N(R^(a))₂,N(R^(a))C(NR^(a))N(R^(a))₂, —N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2),—S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or2), or PO₃(R^(a))₂, where each R^(a) is independently hydrogen, alkyl,fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl,heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

The term “alkoxycarbonyl” refers to a group of the formula(alkoxy)(C═O)— attached through the carbonyl carbon wherein the alkoxygroup has the indicated number of carbon atoms. Thus a(C₁₋₆)alkoxycarbonyl group is an alkoxy group having from 1 to 6 carbonatoms attached through its oxygen to a carbonyl linker. “Loweralkoxycarbonyl” refers to an alkoxycarbonyl group wherein the alkoxygroup is a lower alkoxy group.

The term “substituted alkoxycarbonyl” refers to the group (substitutedalkyl)-O—C(O)— wherein the group is attached to the parent structurethrough the carbonyl functionality. Unless stated otherwise specificallyin the specification, the alkyl moiety of an alkoxycarbonyl group isoptionally substituted by one or more substituents which independentlyare: alkyl, heteroalkyl, acylsulfonamido, alkenyl, alkynyl, cycloalkyl,heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, trimethylsilanyl, —OR^(a), —SR^(a),—S(O)_(t)R^(a)— (where t is 1 or 2), —OC(O)—R^(a), —N(R^(a))₂,—C(O)R^(a), —C(O)OR^(a), —OC(O)N(R^(a))₂, —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a), —N(R^(a))C(O)N(R^(a))₂,N(R^(a))C(NR^(a))N(R^(a))₂, —N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2),—S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or2), or PO₃(R^(a))₂, where each R^(a) is independently hydrogen, alkyl,fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl,heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Acyl” refers to the groups (alkyl)-C(O)—, (aryl)-C(O)—,(heteroaryl)-C(O)—, (heteroalkyl)-C(O)— and (heterocycloalkyl)-C(O)—,wherein the group is attached to the parent structure through thecarbonyl functionality. If the R radical is heteroaryl orheterocycloalkyl, the hetero ring or chain atoms contribute to the totalnumber of chain or ring atoms. Unless stated otherwise specifically inthe specification, the alkyl, aryl or heteroaryl moiety of the acylgroup is optionally substituted by one or more substituents which areindependently alkyl, heteroalkyl, acylsulfonamido, alkenyl, alkynyl,cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, trimethylsilanyl, —OR^(a), —SR^(a),—S(O)_(t)R^(a)— (where t is 1 or 2), —OC(O)—R^(a), —N(R^(a))₂,—C(O)R^(a), —C(O)OR^(a), —OC(O)N(R^(a))₂, —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a), —N(R^(a))C(O)N(R^(a))₂,N(R^(a))C(NR^(a))N(R^(a))₂, —N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2),—S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or2), or PO₃(R^(a))₂, where each R^(a) is independently hydrogen, alkyl,fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl,heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Acyloxy” refers to a R(C═O)O— radical wherein R is alkyl, aryl,heteroaryl, heteroalkyl or heterocycloalkyl, which are as describedherein. If the R radical is heteroaryl or heterocycloalkyl, the heteroring or chain atoms contribute to the total number of chain or ringatoms. Unless stated otherwise specifically in the specification, the Rof an acyloxy group is optionally substituted by one or moresubstituents which independently are: alkyl, heteroalkyl, alkenyl,alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl,heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, trimethylsilanyl, —OR^(a), —SR^(a),—S(O)_(t)R^(a)— (where t is 1 or 2), —OC(O)—R^(a), —N(R^(a))₂,—C(O)R^(a), —C(O)OR^(a), —OC(O)N(R^(a))₂, —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a), —N(R^(a))C(O)N(R^(a))₂,N(R^(a))C(NR^(a))N(R^(a))₂, —N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2),—S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or2), or PO₃(R^(a))₂, where each R^(a) is independently hydrogen, alkyl,fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl,heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Amino” or “amine” refers to a —N(R^(a))₂ radical group, where eachR^(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl, unless statedotherwise specifically in the specification. When a —N(R^(a))₂ group hastwo R^(a) substituents other than hydrogen, they can be combined withthe nitrogen atom to form a 4-, 5-, 6- or 7-membered ring. For example,—N(R^(a))₂ is intended to include, but is not limited to, 1-pyrrolidinyland 4-morpholinyl. Unless stated otherwise specifically in thespecification, an amino group is optionally substituted by one or moresubstituents which independently are: alkyl, acylsulfonamido,heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy,halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl,—OR^(a), —SR^(a), —S(O)_(t)R^(a)— (where t is 1 or 2), —OC(O)—R^(a),—N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —OC(O)N(R^(a))₂, —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a), —N(R^(a))C(O)N(R^(a))₂,N(R^(a))C(NR^(a))N(R^(a))₂, —N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2),—S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or2), or PO₃(R^(a))₂, where each R^(a) is independently hydrogen, alkyl,fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl,heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

The term “substituted amino” also refers to N-oxides of the groups—NHR^(a), and NR^(a)R^(a) each as described above. N-oxides can beprepared by treatment of the corresponding amino group with, forexample, hydrogen peroxide or m-chloroperoxybenzoic acid.

“Amide” or “amido” refers to a chemical moiety with formula —C(O)N(R)₂or —NHC(O)R, where R is selected from the group consisting of hydrogen,alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon) andheteroalicyclic (bonded through a ring carbon), each of which moiety mayitself be optionally substituted. The R₂ of —N(R)₂ of the amide mayoptionally be taken together with the nitrogen to which it is attachedto form a 4-, 5-, 6- or 7-membered ring. Unless stated otherwisespecifically in the specification, an amido group is optionallysubstituted independently by one or more of the substituents asdescribed herein for alkyl, amino, cycloalkyl, aryl, heteroaryl, orheterocycloalkyl. An amide may be an amino acid or a peptide moleculeattached to a compound disclosed herein, thereby forming a prodrug. Theprocedures and specific groups to make such amides are known to those ofskill in the art and can readily be found in seminal sources such asGreene and Wuts, Protective Groups in Organic Synthesis, 3^(rd) Ed.,John Wiley & Sons, New York, N.Y., 1999, which is incorporated herein byreference in its entirety.

“Aromatic” or “aryl” or “Ar” refers to an aromatic radical with six toten ring atoms (e.g., C₆-C₁₀ aromatic or C₆-C₁₀ aryl) which has at leastone ring having a conjugated pi electron system which is carbocyclic(e.g., phenyl, fluorenyl, and naphthyl). Bivalent radicals formed fromsubstituted benzene derivatives and having the free valences at ringatoms are named as substituted phenylene radicals. Bivalent radicalsderived from univalent polycyclic hydrocarbon radicals whose names endin “-yl” by removal of one hydrogen atom from the carbon atom with thefree valence are named by adding “-idene” to the name of thecorresponding univalent radical, e.g., a naphthyl group with two pointsof attachment is termed naphthylidene. Whenever it appears herein, anumerical range such as “6 to 10” refers to each integer in the givenrange; e.g., “6 to 10 ring atoms” means that the aryl group may consistof 6 ring atoms, 7 ring atoms, etc., up to and including 10 ring atoms.The term includes monocyclic or fused-ring polycyclic (i.e., rings whichshare adjacent pairs of ring atoms) groups. Unless stated otherwisespecifically in the specification, an aryl moiety is optionallysubstituted by one or more substituents which are independently alkyl,heteroalkyl, acylsulfonamido, alkenyl, alkynyl, cycloalkyl,heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, trimethylsilanyl, —OR^(a), —SR^(a),—S(O)_(t)R^(a)— (where t is 1 or 2), —OC(O)—R^(a), —N(R^(a))₂,—C(O)R^(a), —C(O)OR^(a), —OC(O)N(R^(a))₂, —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a), —N(R^(a))C(O)N(R^(a))₂,N(R^(a))C(NR^(a))N(R^(a))₂, —N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2),—S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or2), or PO₃(R^(a))₂, where each R^(a) is independently hydrogen, alkyl,fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl,heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Aralkyl” or “arylalkyl” refers to an (aryl)alkyl-radical where aryl andalkyl are as disclosed herein and which are optionally substituted byone or more of the substituents described as suitable substituents foraryl and alkyl respectively.

“Ester” refers to a chemical radical of formula —COOR, where R isselected from the group consisting of alkyl, cycloalkyl, aryl,heteroaryl (bonded through a ring carbon) and heteroalicyclic (bondedthrough a ring carbon). The procedures and specific groups to makeesters are known to those of skill in the art and can readily be foundin seminal sources such as Greene and Wuts, Protective Groups in OrganicSynthesis, 3^(rd) Ed., John Wiley & Sons, New York, N.Y., 1999, which isincorporated herein by reference in its entirety. Unless statedotherwise specifically in the specification, an ester group isoptionally substituted by one or more substituents which independentlyare: alkyl, acylsulfonamido, heteroalkyl, alkenyl, alkynyl, cycloalkyl,heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, trimethylsilanyl, —OR^(a), —SR^(a),—S(O)_(t)R^(a)— (where t is 1 or 2), —OC(O)—R^(a), —N(R^(a))₂,—C(O)R^(a), —C(O)OR^(a), —OC(O)N(R^(a))₂, —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a), —N(R^(a))C(O)N(R^(a))₂,N(R^(a))C(NR^(a))N(R^(a))₂, —N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2),—S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or2), or PO₃(R^(a))₂, where each R^(a) is independently hydrogen, alkyl,fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl,heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Fluoroalkyl” refers to an alkyl radical, as defined above, that issubstituted by one or more fluoro radicals, as defined above, forexample, trifluoromethyl, difluoromethyl, 2,2,2-trifluoroethyl,1-fluoromethyl-2-fluoroethyl, and the like. The alkyl part of thefluoroalkyl radical may be optionally substituted as defined above foran alkyl group.

“Halo,” “halide,” or, alternatively, “halogen” is intended to meanfluoro, chloro, bromo or iodo. The terms “haloalkyl,” “haloalkenyl,”“haloalkynyl,” and “haloalkoxy” include alkyl, alkenyl, alkynyl andalkoxy structures that are substituted with one or more halo groups orwith combinations thereof. For example, the terms “fluoroalkyl” and“fluoroalkoxy” include haloalkyl and haloalkoxy groups, respectively, inwhich the halo is fluorine.

“Heteroalkyl,” “heteroalkenyl,” and “heteroalkynyl” refer to optionallysubstituted alkyl, alkenyl and alkynyl radicals and which have one ormore skeletal chain atoms selected from an atom other than carbon, e.g.,oxygen, nitrogen, sulfur, phosphorus or combinations thereof. Anumerical range may be given—e.g., C₁-C₄ heteroalkyl which refers to thechain length in total, which in this example is 4 atoms long. Aheteroalkyl group may be substituted with one or more substituents whichindependently are: alkyl, heteroalkyl, alkenyl, alkynyl,acylsulfonamido, cycloalkyl, heterocycloalkyl, hydroxamate, aryl,arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, nitro,oxo, thioxo, trimethylsilanyl, —OR^(a), —SR^(a), —S(O)_(t)R^(a)— (wheret is 1 or 2), —OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a),—OC(O)N(R^(a))₂, —C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a),—N(R^(a))C(O)R^(a), —N(R^(a))C(O)N(R^(a))₂, N(R)C(NR^(a))N(R^(a))₂,—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or 2), or PO₃(R^(a))₂, whereeach R^(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Heteroalkylaryl” refers to an -(heteroalkyl)aryl radical whereheteroalkyl and aryl are as disclosed herein and which are optionallysubstituted by one or more of the substituents described as suitablesubstituents for heteroalkyl and aryl, respectively.

“Heteroalkylheteroaryl” refers to an -(heteroalkyl)heteroaryl radicalwhere heteroalkyl and heteroaryl are as disclosed herein and which areoptionally substituted by one or more of the substituents described assuitable substituents for heteroalkyl and heteroaryl, respectively.

“Heteroalkylheterocycloalkyl” refers to an (heteroalkyl)heterocycloalkylradical where heteroalkyl and heterocycloalkyl are as disclosed hereinand which are optionally substituted by one or more of the substituentsdescribed as suitable substituents for heteroalkyl and heterocycloalkyl,respectively.

“Heteroalkylcycloalkyl” refers to an -(heteroalkyl)cycloalkyl radicalwhere heteroalkyl and cycloalkyl are as disclosed herein and which areoptionally substituted by one or more of the substituents described assuitable substituents for heteroalkyl and cycloalkyl, respectively.

“Heteroaryl” or “heteroaromatic” or “HetAr” refers to a 5- to18-membered aromatic radical (e.g., C₅-C₁₃ heteroaryl) that includes oneor more ring heteroatoms selected from nitrogen, oxygen and sulfur, andwhich may be a monocyclic, bicyclic, tricyclic or tetracyclic ringsystem. Whenever it appears herein, a numerical range such as “5 to 18”refers to each integer in the given range—e.g., “5 to 18 ring atoms”means that the heteroaryl group may consist of 5 ring atoms, 6 ringatoms, etc., up to and including 18 ring atoms. Bivalent radicalsderived from univalent heteroaryl radicals whose names end in “-yl” byremoval of one hydrogen atom from the atom with the free valence arenamed by adding “-idene” to the name of the corresponding univalentradical—e.g., a pyridyl group with two points of attachment is apyridylidene. A N-containing “heteroaromatic” or “heteroaryl” moietyrefers to an aromatic group in which at least one of the skeletal atomsof the ring is a nitrogen atom. The polycyclic heteroaryl group may befused or non-fused. The heteroatom(s) in the heteroaryl radical areoptionally oxidized. One or more nitrogen atoms, if present, areoptionally quaternized. The heteroaryl may be attached to the rest ofthe molecule through any atom of the ring(s). Examples of heteroarylsinclude, but are not limited to, azepinyl, acridinyl, benzimidazolyl,benzindolyl, 1,3-benzodioxolyl, benzofuranyl, benzooxazolyl,benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl,benzo[b][1,4]oxazinyl, 1,4-benzodioxanyl, benzonaphthofuranyl,benzoxazolyl, benzodioxolyl, benzodioxinyl, benzoxazolyl, benzopyranyl,benzopyranonyl, benzofuranyl, benzofuranonyl, benzofurazanyl,benzothiazolyl, benzothienyl(benzothiophenyl),benzothieno[3,2-d]pyrimidinyl, benzotriazolyl,benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl,cyclopenta[d]pyrimidinyl,6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl,5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl,6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazinyl, dibenzofuranyl,dibenzothiophenyl, furanyl, furazanyl, furanonyl, furo[3,2-c]pyridinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyridazinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyridinyl, isothiazolyl, imidazolyl,indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl,isoquinolyl, indolizinyl, isoxazolyl, isoxazol-3-one,5,8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl,1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl,5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl,phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl,purinyl, pyranyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl,pyridinyl, pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl,pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl,quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl,5,6,7,8-tetrahydroquinazolinyl,5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl,6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl,5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl,thiapyranyl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl,thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pyridinyl, and thiophenyl (i.e.thienyl). Unless stated otherwise specifically in the specification, aheteroaryl moiety is optionally substituted by one or more substituentswhich are independently: alkyl, acylsulfonamido, heteroalkyl, alkenyl,alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl,heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, nitro, oxo, thioxo,trimethylsilanyl, —OR^(a), —SR^(a), —S(O)_(t)R^(a)— (where t is 1 or 2),—OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —OC(O)N(R^(a))₂,—C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a),—N(R^(a))C(O)N(R^(a))₂, N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or 2), or PO₃(R^(a))₂, whereeach R^(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

Substituted heteroaryl also includes ring systems substituted with oneor more oxide (—O—) substituents, such as, for example, pyridinylN-oxides.

“Heteroarylalkyl” refers to a moiety having an aryl moiety, as describedherein, connected to an alkylene moiety, as described herein, whereinthe connection to the remainder of the molecule is through the alkylenegroup.

“Heterocycloalkyl” refers to a stable 3- to 18-membered non-aromaticring radical that comprises two to twelve carbon atoms and from one tosix heteroatoms selected from nitrogen, oxygen and sulfur. Whenever itappears herein, a numerical range such as “3 to 18” refers to eachinteger in the given range—e.g., “3 to 18 ring atoms” means that theheterocycloalkyl group may consist of 3 ring atoms, 4 ring atoms, etc.,up to and including 18 ring atoms. Unless stated otherwise specificallyin the specification, the heterocycloalkyl radical is a monocyclic,bicyclic, tricyclic or tetracyclic ring system, which may include fusedor bridged ring systems. The heteroatoms in the heterocycloalkyl radicalmay be optionally oxidized. One or more nitrogen atoms, if present, areoptionally quaternized. The heterocycloalkyl radical is partially orfully saturated. The heterocycloalkyl may be attached to the rest of themolecule through any atom of the ring(s). Examples of suchheterocycloalkyl radicals include, but are not limited to, dioxolanyl,thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl,imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl,octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl,2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl,piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl,thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl,thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and1,1-dioxo-thiomorpholinyl. Unless stated otherwise specifically in thespecification, a heterocycloalkyl moiety is optionally substituted byone or more substituents which independently are: alkyl,acylsulfonamido, heteroalkyl, alkenyl, alkynyl, cycloalkyl,heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, nitro, oxo, thioxo,trimethylsilanyl, —OR^(a), —SR^(a), —S(O)_(t)R^(a)— (where t is 1 or 2),—OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —OC(O)N(R^(a))₂,—C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a),—N(R^(a))C(O)N(R^(a))₂, N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or 2), or PO₃(R^(a))₂, whereeach R^(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Heterocycloalkyl” also includes bicyclic ring systems wherein onenon-aromatic ring, usually with 3 to 7 ring atoms, contains at least 2carbon atoms in addition to 1-3 heteroatoms independently selected fromoxygen, sulfur, and nitrogen, as well as combinations comprising atleast one of the foregoing heteroatoms; and the other ring, usually with3 to 7 ring atoms, optionally contains 1-3 heteroatoms independentlyselected from oxygen, sulfur, and nitrogen and is not aromatic.

“Hydroxamate” refers to the —C(O)NR^(a)OR^(a) moiety, where each R^(a)is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Nitro” refers to the —NO₂ radical.

“Oxa” refers to the —O— radical.

“Oxo” refers to the ═O radical.

As used herein, the term “substituted” means that the referenced groupmay have attached one or more additional groups, radicals or moietiesindividually and independently selected from, for example, acyl, alkyl,alkylaryl, cycloalkyl, aralkyl, aryl, carbohydrate, carbonate,heteroaryl, heterocycloalkyl, hydroxamate, hydroxy, alkoxy, aryloxy,mercapto, alkylthio, arylthio, cyano, halo, carbonyl, ester,thiocarbonyl, isocyanato, thiocyanato, isothiocyanato, nitro, oxo,perhaloalkyl, perfluoroalkyl, phosphate, silyl, sulfinyl, sulfonyl,sulfonamidyl, sulfoxyl, sulfonate, urea, and amino, including mono- anddi-substituted amino groups, and protected derivatives thereof. Thesubstituents themselves may be substituted, for example, a cycloalkylsubstituent may itself have a halide substituent at one or more of itsring carbons. The term “optionally substituted” means optionalsubstitution with the specified groups, radicals or moieties. Thechemical moieties of formulas Ia, Ib, II, and/or III, above, that may beoptionally substituted include alkyl, alkenyl, alkynyl, cycloalkyl,arylalkyl, aryl, heterocycle, and heteroaryl. For example, optionallysubstituted alkyl may include both propyl and 2-chloro-propyl.Additionally, “optionally substituted” is also inclusive of embodimentswhere the named substituent or substituents have multiple substituentsrather than simply a single substituent. For example, optionallysubstituted aryl may include both phenyl and3-methyl-5-ethyl-6-chloro-phenyl.

“Isomers” are different compounds that have the same molecular formula.“Stereoisomers” are isomers that differ only in the way the atoms arearranged in space—i.e., having a different stereochemical configuration.“Enantiomers” are a pair of stereoisomers that are non-superimposablemirror images of each other. A 1:1 mixture of a pair of enantiomers is a“racemic” mixture. The term “(±)” is used to designate a racemic mixturewhere appropriate. “Diastereoisomers” are stereoisomers that have atleast two asymmetric atoms, but which are not mirror-images of eachother. The absolute stereochemistry is specified according to theCahn-Ingold-Prelog R-S system. When a compound is a pure enantiomer thestereochemistry at each chiral carbon can be specified by either (R) or(S). Resolved compounds whose absolute configuration is unknown can bedesignated (+) or (−) depending on the direction (dextro- orlevorotatory) which they rotate plane polarized light at the wavelengthof the sodium D line. Certain of the compounds described herein containone or more asymmetric centers and can thus give rise to enantiomers,diastereomers, and other stereoisomeric forms that can be defined, interms of absolute stereochemistry, as (R) or (S). The present chemicalentities, pharmaceutical compositions and methods are meant to includeall such possible isomers, including racemic mixtures, optically pureforms and intermediate mixtures. Optically active (R)- and (S)-isomerscan be prepared using chiral synthons or chiral reagents, or resolvedusing conventional techniques. When the compounds described hereincontain olefinic double bonds or other centers of geometric asymmetry,and unless specified otherwise, it is intended that the compoundsinclude both E and Z geometric isomers.

“Enantiomeric purity” as used herein refers to the relative amounts,expressed as a percentage, of the presence of a specific enantiomerrelative to the other enantiomer. For example, if a compound, which maypotentially have an (R)- or an (S)-isomeric configuration, is present asa racemic mixture, the enantiomeric purity is about 50% with respect toeither the (R)- or (S)-isomer. If that compound has one isomeric formpredominant over the other, for example, 80% (S)-isomer and 20%(R)-isomer, the enantiomeric purity of the compound with respect to the(S)-isomeric form is 80%. The enantiomeric purity of a compound can bedetermined in a number of ways known in the art, including but notlimited to chromatography using a chiral support, polarimetricmeasurement of the rotation of polarized light, nuclear magneticresonance spectroscopy using chiral shift reagents which include but arenot limited to lanthanide containing chiral complexes or Pirkle'sreagents, or derivatization of a compounds using a chiral compound suchas Mosher's acid followed by chromatography or nuclear magneticresonance spectroscopy.

In some embodiments, the enantiomerically enriched composition has ahigher potency with respect to therapeutic utility per unit mass thandoes the racemic mixture of that composition. Enantiomers can beisolated from mixtures by methods known to those skilled in the art,including chiral high pressure liquid chromatography (HPLC) and theformation and crystallization of chiral salts; or preferred enantiomerscan be prepared by asymmetric syntheses. See, for example, Jacques, etal., Enantiomers, Racemates and Resolutions, Wiley Interscience, NewYork (1981); E. L. Eliel, Stereochemistry of Carbon Compounds,McGraw-Hill, New York (1962); and E. L. Eliel and S. H. Wilen,Stereochemistry of Organic Compounds, Wiley-Interscience, New York(1994).

The terms “enantiomerically enriched” and “non-racemic,” as used herein,refer to compositions in which the percent by weight of one enantiomeris greater than the amount of that one enantiomer in a control mixtureof the racemic composition (e.g., greater than 1:1 by weight). Forexample, an enantiomerically enriched preparation of the (S)-enantiomer,means a preparation of the compound having greater than 50% by weight ofthe (S)-enantiomer relative to the (R)-enantiomer, such as at least 75%by weight, or such as at least 80% by weight. In some embodiments, theenrichment can be significantly greater than 80% by weight, providing a“substantially enantiomerically enriched” or a “substantiallynon-racemic” preparation, which refers to preparations of compositionswhich have at least 85% by weight of one enantiomer relative to otherenantiomer, such as at least 90% by weight, or such as at least 95% byweight. The terms “enantiomerically pure” or “substantiallyenantiomerically pure” refers to a composition that comprises at least98% of a single enantiomer and less than 2% of the opposite enantiomer.

“Moiety” refers to a specific segment or functional group of a molecule.Chemical moieties are often recognized chemical entities embedded in orappended to a molecule.

“Tautomers” are structurally distinct isomers that interconvert bytautomerization. “Tautomerization” is a form of isomerization andincludes prototropic or proton-shift tautomerization, which isconsidered a subset of acid-base chemistry. “Prototropictautomerization” or “proton-shift tautomerization” involves themigration of a proton accompanied by changes in bond order, often theinterchange of a single bond with an adjacent double bond. Wheretautomerization is possible (e.g., in solution), a chemical equilibriumof tautomers can be reached. An example of tautomerization is keto-enoltautomerization. A specific example of keto-enol tautomerization is theinterconversion of pentane-2,4-dione and 4-hydroxypent-3-en-2-onetautomers. Another example of tautomerization is phenol-ketotautomerization. A specific example of phenol-keto tautomerization isthe interconversion of pyridin-4-ol and pyridin-4(1H)-one tautomers.

A “leaving group or atom” is any group or atom that will, under selectedreaction conditions, cleave from the starting material, thus promotingreaction at a specified site. Examples of such groups, unless otherwisespecified, include halogen atoms and mesyloxy, p-nitrobenzensulphonyloxyand tosyloxy groups.

“Protecting group” is intended to mean a group that selectively blocksone or more reactive sites in a multifunctional compound such that achemical reaction can be carried out selectively on another unprotectedreactive site and the group can then be readily removed or deprotectedafter the selective reaction is complete. A variety of protecting groupsare disclosed, for example, in T. H. Greene and P. G. M. Wuts,Protective Groups in Organic Synthesis, 3rd Edition, John Wiley & Sons,New York (1999).

“Solvate” refers to a compound in physical association with one or moremolecules of a pharmaceutically acceptable solvent.

Compounds of the invention also include crystalline and amorphous formsof those compounds, including, for example, polymorphs,pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (includinganhydrates), conformational polymorphs, and amorphous forms of thecompounds, as well as mixtures thereof “Crystalline form” and“polymorph” are intended to include all crystalline and amorphous formsof the compound, including, for example, polymorphs, pseudopolymorphs,solvates, hydrates, unsolvated polymorphs (including anhydrates),conformational polymorphs, and amorphous forms, as well as mixturesthereof, unless a particular crystalline or amorphous form is referredto.

Compounds of the invention also include antibodies. The terms “antibody”and its plural form “antibodies” refer to whole immunoglobulins and anyantigen-binding fragment (“antigen-binding portion”) or single chainsthereof. An “antibody” further refers to a glycoprotein comprising atleast two heavy (H) chains and two light (L) chains inter-connected bydisulfide bonds, or an antigen-binding portion thereof. Each heavy chainis comprised of a heavy chain variable region (abbreviated herein asV_(H)) and a heavy chain constant region. The heavy chain constantregion is comprised of three domains, CH1, CH2 and CH3. Each light chainis comprised of a light chain variable region (abbreviated herein asV_(L)) and a light chain constant region. The light chain constantregion is comprised of one domain, C_(L). The V_(H) and V_(L) regions ofan antibody may be further subdivided into regions of hypervariability,which are referred to as complementarity determining regions (CDR) orhypervariable regions (HVR), and which can be interspersed with regionsthat are more conserved, termed framework regions (FR). Each V_(H) andV_(L) is composed of three CDRs and four FRs, arranged fromamino-terminus to carboxy-terminus in the following order: FR1, CDR1,FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and lightchains contain a binding domain that interacts with an antigen epitopeor epitopes. The constant regions of the antibodies may mediate thebinding of the immunoglobulin to host tissues or factors, includingvarious cells of the immune system (e.g., effector cells) and the firstcomponent (Clq) of the classical complement system.

The terms “monoclonal antibody,” “mAb,” “monoclonal antibodycomposition,” or their plural forms refer to a preparation of antibodymolecules of single molecular composition. A monoclonal antibodycomposition displays a single binding specificity and affinity for aparticular epitope. Monoclonal antibodies recited herein and/or thosespecific to, e.g., PD-1 or PD-L1, can be made using knowledge and skillin the art of injecting test subjects with a corresponding antigen andthen isolating hybridomas expressing antibodies having the desiredsequence or functional characteristics. DNA encoding the monoclonalantibodies is readily isolated and sequenced using conventionalprocedures (e.g., by using oligonucleotide probes that are capable ofbinding specifically to genes encoding the heavy and light chains of themonoclonal antibodies). The hybridoma cells serve as a preferred sourceof such DNA. Once isolated, the DNA may be placed into expressionvectors, which are then transfected into host cells such as E. colicells, simian COS cells, Chinese hamster ovary (CHO) cells, or myelomacells that do not otherwise produce immunoglobulin protein, to obtainthe synthesis of monoclonal antibodies in the recombinant host cells.Recombinant production of antibodies will be described in more detailbelow.

The terms “antigen-binding portion” or “antigen-binding fragment” of anantibody (or simply “antibody portion”), as used herein, refers to oneor more fragments of an antibody that retain the ability to specificallybind to an antigen. It has been shown that the antigen-binding functionof an antibody can be performed by fragments of a full-length antibody.Examples of binding fragments encompassed within the term“antigen-binding portion” of an antibody include (i) a Fab fragment, amonovalent fragment consisting of the V_(L), V_(H), C_(L) and CH1domains; (ii) a F(ab′)2 fragment, a bivalent fragment comprising two Fabfragments linked by a disulfide bridge at the hinge region; (iii) a Fdfragment consisting of the V_(H) and CH1 domains; (iv) a Fv fragmentconsisting of the V_(L) and V_(H) domains of a single arm of anantibody, (v) a domain antibody (dAb) fragment (Ward et al., Nature,1989, 341, 544-546), which may consist of a V_(H) or a V_(L) domain; and(vi) an isolated complementarity determining region (CDR). Furthermore,although the two domains of the Fv fragment, V_(L) and V_(H), are codedfor by separate genes, they can be joined, using recombinant methods, bya synthetic linker that enables them to be made as a single proteinchain in which the V_(L) and V_(H) regions pair to form monovalentmolecules known as single chain Fv (scFv); see, e.g., Bird et al.,Science 1988, 242, 423-426; and Huston et al., Proc. Natl. Acad. Sci.USA 1988, 85, 5879-5883). Such scFv antibodies are also intended to beencompassed within the terms “antigen-binding portion” or“antigen-binding fragment” of an antibody. These antibody fragments areobtained using conventional techniques known to those with skill in theart, and the fragments are screened for utility in the same manner asare intact antibodies.

The term “human antibody,” as used herein, is intended to includeantibodies having variable regions in which both the framework and CDRregions are derived from human germline immunoglobulin sequences.Furthermore, if the antibody contains a constant region, the constantregion also is derived from human germline immunoglobulin sequences. Thehuman antibodies of the invention may include amino acid residues notencoded by human germline immunoglobulin sequences (e.g., mutationsintroduced by random or site-specific mutagenesis in vitro or by somaticmutation in vivo). The term “human antibody”, as used herein, is notintended to include antibodies in which CDR sequences derived from thegermline of another mammalian species, such as a mouse, have beengrafted onto human framework sequences.

The term “human monoclonal antibody” refers to antibodies displaying asingle binding specificity which have variable regions in which both theframework and CDR regions are derived from human germline immunoglobulinsequences. In one embodiment, the human monoclonal antibodies areproduced by a hybridoma which includes a B cell obtained from atransgenic nonhuman animal, e.g., a transgenic mouse, having a genomecomprising a human heavy chain transgene and a light chain transgenefused to an immortalized cell.

The term “recombinant human antibody”, as used herein, includes allhuman antibodies that are prepared, expressed, created or isolated byrecombinant means, such as (a) antibodies isolated from an animal (e.g.,a mouse) that is transgenic or transchromosomal for human immunoglobulingenes or a hybridoma prepared therefrom (described further below), (b)antibodies isolated from a host cell transformed to express the humanantibody, e.g., from a transfectoma, (c) antibodies isolated from arecombinant, combinatorial human antibody library, and (d) antibodiesprepared, expressed, created or isolated by any other means that involvesplicing of human immunoglobulin gene sequences to other DNA sequences.Such recombinant human antibodies have variable regions in which theframework and CDR regions are derived from human germline immunoglobulinsequences. In certain embodiments, however, such recombinant humanantibodies can be subjected to in vitro mutagenesis (or, when an animaltransgenic for human Ig sequences is used, in vivo somatic mutagenesis)and thus the amino acid sequences of the V_(H) and V_(L) regions of therecombinant antibodies are sequences that, while derived from andrelated to human germline V_(H) and V_(L) sequences, may not naturallyexist within the human antibody germline repertoire in vivo.

As used herein, “isotype” refers to the antibody class (e.g., IgM orIgG1) that is encoded by the heavy chain constant region genes. Inmammals, there are five antibody isotypes: IgA, IgD, IgG, IgM and IgE.In humans, there are four subclasses of the IgG isotype: IgG1, IgG2,IgG3 and IgG4, and two subclasses of the IgA isotype: IgA1 and IgA2.

The phrases “an antibody recognizing an antigen” and “an antibodyspecific for an antigen” are used interchangeably herein with the term“an antibody which binds specifically to an antigen.”

The term “human antibody derivatives” refers to any modified form of thehuman antibody, e.g., a conjugate of the antibody and another activepharmaceutical ingredient or antibody. The terms “conjugate,”“antibody-drug conjugate”, “ADC,” or “immunoconjugate” refers to anantibody, or a fragment thereof, conjugated to a therapeutic moiety,such as a bacterial toxin, a cytotoxic drug or a radionuclide-containingtoxin. Toxic moieties can be conjugated to antibodies of the inventionusing methods available in the art.

The terms “humanized antibody,” “humanized antibodies,” and “humanized”are intended to refer to antibodies in which CDR sequences derived fromthe germline of another mammalian species, such as a mouse, have beengrafted onto human framework sequences. Additional framework regionmodifications may be made within the human framework sequences.Humanized forms of non-human (for example, murine) antibodies arechimeric antibodies that contain minimal sequence derived from non-humanimmunoglobulin. For the most part, humanized antibodies are humanimmunoglobulins (recipient antibody) in which residues from ahypervariable region of the recipient are replaced by residues from a 15hypervariable region of a non-human species (donor antibody) such asmouse, rat, rabbit or nonhuman primate having the desired specificity,affinity, and capacity. In some instances, Fv framework region (FR)residues of the human immunoglobulin are replaced by correspondingnon-human residues. Furthermore, humanized antibodies may compriseresidues that are not found in the recipient antibody or in the donorantibody. These modifications are made to further refine antibodyperformance. In general, the humanized antibody will comprisesubstantially all of at least one, and typically two, variable domains,in which all or substantially all of the hypervariable loops correspondto those of a non-human immunoglobulin and all or substantially all ofthe FR regions are those of a human immunoglobulin sequence. Thehumanized antibody optionally also will comprise at least a portion ofan immunoglobulin constant region (Fc), typically that of a humanimmunoglobulin. For further details, see Jones et al., Nature 1986, 321,522-525; Riechmann et al., Nature 1988, 332, 323-329; and Presta, Curr.Op. Struct. Biol. 1992, 2, 593-596.

The term “chimeric antibody” is intended to refer to antibodies in whichthe variable region sequences are derived from one species and theconstant region sequences are derived from another species, such as anantibody in which the variable region sequences are derived from a mouseantibody and the constant region sequences are derived from a humanantibody.

A “diabody” is a small antibody fragment with two antigen-binding sites.The fragments comprise a heavy chain variable domain (V_(H)) connectedto a light chain variable domain (V_(L)) in the same polypeptide chain(V_(H)-V_(L) or V_(L)-V_(H)). By using a linker that is too short toallow pairing between the two domains on the same chain, the domains areforced to pair with the complementary domains of another chain andcreate two antigen-binding sites. Diabodies are described more fully in,e.g., European Patent No. EP 404,097, International Patent PublicationNo. WO 93/11161; and Bolliger et al., Proc. Natl. Acad. Sci. USA 1993,90, 6444-6448.

The term “glycosylation” refers to a modified derivative of an antibody.An aglycoslated antibody lacks glycosylation. Glycosylation can bealtered to, for example, increase the affinity of the antibody forantigen. Such carbohydrate modifications can be accomplished by, forexample, altering one or more sites of glycosylation within the antibodysequence. For example, one or more amino acid substitutions can be madethat result in elimination of one or more variable region frameworkglycosylation sites to thereby eliminate glycosylation at that site.Aglycosylation may increase the affinity of the antibody for antigen, asdescribed in U.S. Pat. Nos. 5,714,350 and 6,350,861. Additionally oralternatively, an antibody can be made that has an altered type ofglycosylation, such as a hypofucosylated antibody having reduced amountsof fucosyl residues or an antibody having increased bisecting GlcNacstructures. Such altered glycosylation patterns have been demonstratedto increase the ability of antibodies. Such carbohydrate modificationscan be accomplished by, for example, expressing the antibody in a hostcell with altered glycosylation machinery. Cells with alteredglycosylation machinery have been described in the art and can be usedas host cells in which to express recombinant antibodies of theinvention to thereby produce an antibody with altered glycosylation. Forexample, the cell lines Ms704, Ms705, and Ms709 lack thefucosyltransferase gene, FUT8 (alpha (1,6) fucosyltransferase), suchthat antibodies expressed in the Ms704, Ms705, and Ms709 cell lines lackfucose on their carbohydrates. The Ms704, Ms705, and Ms709 FUT8−/− celllines were created by the targeted disruption of the FUT8 gene inCHO/DG44 cells using two replacement vectors (see e.g. U.S. PatentPublication No. 2004/0110704 or Yamane-Ohnuki, et al. Biotechnol.Bioeng., 2004, 87, 614-622). As another example, European Patent No. EP1,176,195 describes a cell line with a functionally disrupted FUT8 gene,which encodes a fucosyl transferase, such that antibodies expressed insuch a cell line exhibit hypofucosylation by reducing or eliminating thealpha 1,6 bond-related enzyme, and also describes cell lines which havea low enzyme activity for adding fucose to the N-acetylglucosamine thatbinds to the Fc region of the antibody or does not have the enzymeactivity, for example the rat myeloma cell line YB2/0 (ATCC CRL 1662).International Patent Publication WO 03/035835 describes a variant CHOcell line, Lec 13 cells, with reduced ability to attach fucose toAsn(297)-linked carbohydrates, also resulting in hypofucosylation ofantibodies expressed in that host cell (see also Shields, et al., J.Biol. Chem. 2002, 277, 26733-26740. International Patent Publication WO99/54342 describes cell lines engineered to expressglycoprotein-modifying glycosyl transferases (e.g.,beta(1,4)-N-acetylglucosaminyltransferase III (GnTIII)) such thatantibodies expressed in the engineered cell lines exhibit increasedbisecting GlcNac structures which results in increased ADCC activity ofthe antibodies (see also Umana, et al., Nat. Biotech. 1999, 17,176-180). Alternatively, the fucose residues of the antibody may becleaved off using a fucosidase enzyme. For example, the fucosidasealpha-L-fucosidase removes fucosyl residues from antibodies as describedin Tarentino, et al., Biochem. 1975, 14, 5516-5523.

“Pegylation” refers to a modified antibody, or a fragment thereof, thattypically is reacted with polyethylene glycol (PEG), such as a reactiveester or aldehyde derivative of PEG, under conditions in which one ormore PEG groups become attached to the antibody or antibody fragment.Pegylation may, for example, increase the biological (e.g., serum) halflife of the antibody. Preferably, the pegylation is carried out via anacylation reaction or an alkylation reaction with a reactive PEGmolecule (or an analogous reactive water-soluble polymer). As usedherein, the term “polyethylene glycol” is intended to encompass any ofthe forms of PEG that have been used to derivatize other proteins, suchas mono (C₁-C₁₀) alkoxy- or aryloxy-polyethylene glycol or polyethyleneglycol-maleimide. The antibody to be pegylated may be an aglycosylatedantibody. Methods for pegylation are known in the art and can be appliedto the antibodies of the invention, as described for example in EuropeanPatent Nos. EP 0154316 and EP 0401384.

The term “radioisotope-labeled complex” refers to both non-covalent andcovalent attachment of a radioactive isotope, such as ⁹⁰Y, ¹¹¹In, or¹³¹I, to an antibody, including conjugates.

The term “biosimilar” means a biological product that is highly similarto a U.S. licensed reference biological product notwithstanding minordifferences in clinically inactive components, and for which there areno clinically meaningful differences between the biological product andthe reference product in terms of the safety, purity, and potency of theproduct. Furthermore, a similar biological or “biosimilar” medicine is abiological medicine that is similar to another biological medicine thathas already been authorized for use by the European Medicines Agency.The term “biosimilar” is also used synonymously by other national andregional regulatory agencies. Biological products or biologicalmedicines are medicines that are made by or derived from a biologicalsource, such as a bacterium or yeast. They can consist of relativelysmall molecules such as human insulin or erythropoietin, or complexmolecules such as monoclonal antibodies. For example, if the referenceanti-CD20 monoclonal antibody is rituximab, an anti-CD20 biosimilarmonoclonal antibody approved by drug regulatory authorities withreference to rituximab is a “biosimilar to” rituximab or is a“biosimilar thereof” of rituximab. In Europe, a similar biological or“biosimilar” medicine is a biological medicine that is similar toanother biological medicine that has already been authorized for use bythe European Medicines Agency (EMA). The relevant legal basis forsimilar biological applications in Europe is Article 6 of Regulation(EC) No 726/2004 and Article 10(4) of Directive 2001/83/EC, as amendedand therefore in Europe, the biosimilar may be authorised, approved forauthorisation or subject of an application for authorisation underArticle 6 of Regulation (EC) No 726/2004 and Article 10(4) of Directive2001/83/EC. The already authorized original biological medicinal productmay be referred to as a “reference medicinal product” in Europe. Some ofthe requirements for a product to be considered a biosimilar areoutlined in the CHMP Guideline on Similar Biological Medicinal Products.In addition, product specific guidelines, including guidelines relatingto monoclonal antibody biosimilars, are provided on a product-by-productbasis by the EMA and published on its website. A biosimilar as describedherein may be similar to the reference medicinal product by way ofquality characteristics, biological activity, mechanism of action,safety profiles and/or efficacy. In addition, the biosimilar may be usedor be intended for use to treat the same conditions as the referencemedicinal product. Thus, a biosimilar as described herein may be deemedto have similar or highly similar quality characteristics to a referencemedicinal product. Alternatively, or in addition, a biosimilar asdescribed herein may be deemed to have similar or highly similarbiological activity to a reference medicinal product. Alternatively, orin addition, a biosimilar as described herein may be deemed to have asimilar or highly similar safety profile to a reference medicinalproduct. Alternatively, or in addition, a biosimilar as described hereinmay be deemed to have similar or highly similar efficacy to a referencemedicinal product. As described herein, a biosimilar in Europe iscompared to a reference medicinal product which has been authorised bythe EMA. However, in some instances, the biosimilar may be compared to abiological medicinal product which has been authorised outside theEuropean Economic Area (a non-EEA authorised “comparator”) in certainstudies. Such studies include for example certain clinical and in vivonon-clinical studies. As used herein, the term “biosimilar” also relatesto a biological medicinal product which has been or may be compared to anon-EEA authorised comparator. Certain biosimilars are proteins such asantibodies, antibody fragments (for example, antigen binding portions)and fusion proteins. A protein biosimilar may have an amino acidsequence that has minor modifications in the amino acid structure(including for example deletions, additions, and/or substitutions ofamino acids) which do not significantly affect the function of thepolypeptide. The biosimilar may comprise an amino acid sequence having asequence identity of 97% or greater to the amino acid sequence of itsreference medicinal product, e.g., 97%, 98%, 99% or 100%. The biosimilarmay comprise one or more post-translational modifications, for example,although not limited to, glycosylation, oxidation, deamidation, and/ortruncation which is/are different to the post-translationalmodifications of the reference medicinal product, provided that thedifferences do not result in a change in safety and/or efficacy of themedicinal product. The biosimilar may have an identical or differentglycosylation pattern to the reference medicinal product. Particularly,although not exclusively, the biosimilar may have a differentglycosylation pattern if the differences address or are intended toaddress safety concerns associated with the reference medicinal product.Additionally, the biosimilar may deviate from the reference medicinalproduct in for example its strength, pharmaceutical form, formulation,excipients and/or presentation, providing safety and efficacy of themedicinal product is not compromised. The biosimilar may comprisedifferences in for example pharmacokinetic (PK) and/or pharmacodynamic(PD) profiles as compared to the reference medicinal product but isstill deemed sufficiently similar to the reference medicinal product asto be authorised or considered suitable for authorisation. In certaincircumstances, the biosimilar exhibits different binding characteristicsas compared to the reference medicinal product, wherein the differentbinding characteristics are considered by a Regulatory Authority such asthe EMA not to be a barrier for authorisation as a similar biologicalproduct. The term “biosimilar” is also used synonymously by othernational and regional regulatory agencies.

The terms “sequence identity,” “percent identity,” and “sequence percentidentity” in the context of two or more nucleic acids or polypeptides,refer to two or more sequences or subsequences that are the same or havea specified percentage of nucleotides or amino acid residues that arethe same, when compared and aligned (introducing gaps, if necessary) formaximum correspondence, not considering any conservative amino acidsubstitutions as part of the sequence identity. The percent identity canbe measured using sequence comparison software or algorithms or byvisual inspection. Various algorithms and software are known in the artthat can be used to obtain alignments of amino acid or nucleotidesequences. Suitable programs to determine percent sequence identityinclude for example the BLAST suite of programs available from the U.S.Government's National Center for Biotechnology Information BLAST website. Comparisons between two sequences can be carried using either theBLASTN or BLASTP algorithm. BLASTN is used to compare nucleic acidsequences, while BLASTP is used to compare amino acid sequences. ALIGN,ALIGN-2 (Genentech, South San Francisco, Calif.) or MegAlign, availablefrom DNASTAR, are additional publicly available software programs thatcan be used to align sequences. One skilled in the art can determineappropriate parameters for maximal alignment by particular alignmentsoftware. In certain embodiments, the default parameters of thealignment software are used.

Certain embodiments described herein may comprise a variant of anantibody. As used herein, the term “variant” encompasses but is notlimited to antibodies which comprise an amino acid sequence whichdiffers from the amino acid sequence of a reference antibody by way ofone or more substitutions, deletions and/or additions at certainpositions within or adjacent to the amino acid sequence of the referenceantibody. The variant may comprise one or more conservativesubstitutions in its amino acid sequence as compared to the amino acidsequence of a reference antibody. Conservative substitutions mayinvolve, e.g., the substitution of similarly charged or uncharged aminoacids. The variant retains the ability to specifically bind to theantigen of the reference antibody.

For the avoidance of doubt, it is intended herein that particularfeatures (for example integers, characteristics, values, uses, diseases,formulae, compounds or groups) described in conjunction with aparticular aspect, embodiment or example of the invention are to beunderstood as applicable to any other aspect, embodiment or exampledescribed herein unless incompatible therewith. Thus such features maybe used where appropriate in conjunction with any of the definition,claims or embodiments defined herein. All of the features disclosed inthis specification (including any accompanying claims, abstract anddrawings), and/or all of the steps of any method or process sodisclosed, may be combined in any combination, except combinations whereat least some of the features and/or steps are mutually exclusive. Theinvention is not restricted to any details of any disclosed embodiments.The invention extends to any novel one, or novel combination, of thefeatures disclosed in this specification (including any accompanyingclaims, abstract and drawings), or to any novel one, or any novelcombination, of the steps of any method or process so disclosed.

Histone Deacetylase (HDAC) Protein

Histone acetylation and deacetylation play important roles in chromatinfolding and maintenance. Acetylation appears to play a role in theepigenetic regulation of chromatin structure, and gene expression,through the balance of histone acetyltransferase (HAT) and histonedeacetylase (HDAC) activities. Increased acetylation of histones leadsto changes in chromatin structure and accessibility for key cellularproteins to specific target sites. HATs acetylate lysine groups at theamino terminal tails of nuclear histones to neutralize positive chargeson the histones, yielding a more open, transcriptionally activechromatin structure. In contrast, the HDACs deacetylate and suppresstranscription. In this model, inhibitors of HDACs bias the balancetoward a more acetylated state. Such a shift in the relative activitiesof these enzymes may affect gene expression necessary for DNA repair,replication, cell cycle checkpoint activation and tumor suppression.

Human HDACs may be divided into four classes based on structure,sequence homology, and domain organization. Class I consists of HDACs 1,2, 3, 8, and 11, albeit a recent report puts HDAC 11 into a new class,class IV, based on a phylogenetic analysis. Class I HDACs are nuclearand play roles in cell proliferation and apoptosis. Class II includesHDACs 4, 5, 6, 7, 9, and 10. These enzymes are characterized by a largeNH₂-terminal domain or a second catalytic site and their expression ismore restricted, suggesting roles in cellular differentiation anddevelopment. Class III enzymes, include the sirtuins (SIRTs), and areNAD-dependent deacetylases. These are not inhibited by Trichostatin A(TSA) or other hydroxamates.

HDACs are found in the nuclear and cytoplasmic compartments. Althoughthey are involved in critical cellular functions, such as cell cycleregulation and apoptosis, a key function of HDACs is transcriptionalregulation. HDACs function as components of large multi-proteincomplexes that bind to promoters and repress transcription. Class IIcompounds shuttle between the nucleus and the cytoplasm. However,certain classes of HDACs have conserved deacetylase core domains ofapproximately 400 amino acids and zinc binding sites. It is the coredomain that presents the principal target for design of inhibitory smallmolecules.

In response to DNA damage, signal transduction pathways may be activatedto regulate cell cycle arrest, repair, differentiation, apoptosis, andtranscription. Such responses are a complex feature of the cellularradiation phenotype, and their effectiveness may determine cell survivalor death. DNA damage checkpoints generate signals that arrest cell cycleprogression until the damage is repaired. When damaged DNA is repaired,checkpoint signals are reversed to resume cell cycle progression. SuchDNA-directed processes are accompanied by highly localized changes inchromatin structure. Various recent studies have implicated chromatinstructure in DNA damage signaling and repair. Post-translational histonemodifications regulate chromatin structure and access for proteins todamaged DNA sites as reported for repair and signaling proteins to thedamaged regions of DNA.

Early HDAC inhibitors (e.g., benzamides) were investigated asdifferentiating agents, without full understanding of their molecularmechanisms. Some of these agents have advanced to clinical trials. Thefull recognition of the potential for HDAC inhibitors was advanced withthe discovery and development of hydroxamic acid inhibitors. Hydroxamicacid based compounds (e.g., suberoylanilide hydroxamic acid (SAHA)) havebeen developed for clinical application, and have proven to berelatively non-toxic. SAHA has been approved by the FDA for thetreatment of cutaneous T-cell lymphoma. Certain HDAC inhibitors havebeen described in U.S. Pat. Nos. 7,507,828; 7,842,835; 8,067,600;8,222,451; and 8,748,463; the entirety of which are incorporated hereinby reference.

Other chemical families of HDAC inhibitors, including depsipeptide andvalproic acid, have been shown to inhibit cancer cell growth in vitroand in vivo. Modulation of p53, ErbB1, ErbB2 and Raf-1 expression havebeen observed following exposure of lung cancer cells to depsipeptide, adrug currently in clinical trials. For example, Valproic acid has beenused clinically as an anti-epileptic agent, with excellent reasonabletoxicity profile and has been shown to be involved in the proteolysis ofHDAC2.

However, pan-HDAC inhibitors have multiple toxicities due to off-targeteffects. A molecule targeting the HDAC6 protein has a better chance fortreating diseases, as compared to more promiscuous HDAC inhibitors, withless toxicity.

HDAC Inhibitors

The HDAC inhibitors of the invention may include one or more compoundsof formulas I, II, III, IV, V, VI, VII, and/or VIII. In someembodiments, the HDAC inhibitors of the invention are selective HDAC6inhibitors.

In some embodiments, the HDAC inhibitor of the invention is a compoundof formula VI or a pharmaceutically acceptable salt, solvate, hydrate,cocrystal, or prodrug thereof.

In some embodiments, the HDAC inhibitor of the invention is a compoundof formula VII or a pharmaceutically acceptable salt, solvate, hydrate,cocrystal, or prodrug thereof.

In some embodiments, the HDAC inhibitor of the invention is a compoundof formula VIII or a pharmaceutically acceptable salt, solvate, hydrate,cocrystal, or prodrug thereof.

Certain HDAC inhibitors of the invention are selective HDAC6 inhibitors.For example, the compounds of formulas VI, VII, and VIII are HDACinhibitors that are also selective HDAC6 inhibitors.

In some embodiments, the HDAC inhibitors (e.g., selective HDAC6inhibitors) described herein may be delivered as listed or as apharmaceutically acceptable salts, solvates, hydrates, cocrystals,tautomers, or prodrugs thereof.

In some embodiments, the pharmaceutical compositions and methodsdescribed herein include one or more of the foregoing HDAC inhibitors orselective HDAC6 inhibitors.

Chemotherapeutic Agents

In some embodiments, the pharmaceutical compositions and methodsdescribed herein may include one or more additional therapeutic agents.The additional therapeutic agents may include one or morechemotherapeutic agents.

In some embodiments, the chemotherapeutic agents may include one or moreof bortezomib (Velcade®; see, e.g., U.S. Pat. Nos. 5,780,454, 6,713,446,and 6,958,319), lenalidomide (Revlimid®; see, e.g., U.S. Pat. Nos.5,635,517, 6,045,501, 6,281,230, 6,315,720, 6,555,554, 6,561,976,6,561,976, 6,561,977, 6,755,784, 6,908,432, 7,119,106, 7,189,740,7,465,800, 7,468,363, 7,855,217, 7,968,569, 8,204,763, 8,288,415,8,315,886, 8,404,717, 8,530,498, 8,589,188, 8,626,531, 8,648,095,8,741,929, 9,056,120, 9,101,621, and 9,101,622), temozolomide (see,e.g., U.S. Pat. No. 5,939,098), 5-fluorouracil (5-FU; see, e.g., U.S.Pat. No. 4,130,648), 6-mercaptopurine (6-MP; see, e.g., U.S. Pat. No.8,188,067), Capecitabine (Xeloda®; see, e.g., U.S. Pat. Nos. 4,966,891,5,453,497, 5,472,949, and 5,476,932), Cladribine (see, e.g., U.S. Pat.No. 6,271,245), Clofarabine (see, e.g., U.S. Pat. No. 5,661,136),Cytarabine (Ara-C®; see, e.g., U.S. Pat. Nos. 5,455,044 and 5,723,147),Floxuridine (see, e.g., U.S. Pat. No. 6,271,245), Fludarabine (see,e.g., U.S. Pat. No. 6,271,245), Gemcitabine (Gemzar®; see, e.g., U.S.Pat. Nos. 4,808,614 and 5,464,826), Hydroxyurea (see, U.S. Pat. No.3,968,249), Methotrexate (see, e.g., U.S. Pat. No. 6,271,245),Pemetrexed (Alimta®; see, e.g., U.S. Pat. Nos. 5,344,932 and 7,772,209),Pentostatin (see, e.g., U.S. Pat. No. 6,316,435), Thioguanine (see,e.g., U.S. Pat. No. 3,163,639), daunorubicin (see, e.g., U.S. Pat. No.7,727,968), doxorubicin (see, e.g., U.S. Pat. No. 6,271,245), epirubicin(see, e.g., U.S. Pat. No. 8,410,131), idaurubicin (see, e.g., U.S. Pat.No. 6,271,245), topotecan (see, e.g., U.S. Pat. No. 8,410,131),irinotecan (see, e.g., U.S. Pat. No. 8,410,131), etoposide (see, e.g.,U.S. Pat. No. 6,271,245), teniposide (see, e.g., U.S. Pat. No.6,271,245), mitoxantrone (see, e.g., U.S. Pat. No. 6,271,245),Vinblastine (see, e.g., U.S. Pat. No. 6,271,245), vincristine (see,e.g., U.S. Pat. No. 6,271,245), vinorelbine (see, e.g., U.S. Pat. No.8,410,131), estramustine (see, e.g., U.S. Pat. No. 6,849,616),paclitaxel (see, e.g., U.S. Pat. No. 6,271,245), dexamethasone (see,e.g., U.S. Pat. No. 8,822,438), and docetaxel (see, e.g., U.S. Pat. No.8,410,131).

In any of the methods described herein, the chemotherapeutic agents orcombinations thereof may be administered before, concurrently, or afteradministration of the compounds of formulas I, II, III, IV, V, VI, VII,and/or VIII and/or the immunotherapeutic agents described herein.

Immunotherapeutic Agents

In some embodiments, the pharmaceutical compositions and methodsdescribed herein may include one or more additional therapeutic agents.The additional therapeutic agents may include one or moreimmunotherapeutic agents.

In some embodiments, the immunotherapeutic agents may include one ormore of a PD-1 inhibitor and a PD-L1 inhibitor, wherein the PD-1inhibitor and PD-L1 inhibitor may be selected from the group consistingof nivolumab, pembrolizumab, pidilizumab, durvalumab, atezolizumab, andavelumab, and any fragment, derivative, conjugate, variant,radioisotope-labeled complex, or biosimilar thereof.

In some embodiments, the immunotherapeutic agents may include one ormore of rituximab, trastuzumab, ibritumomab, tositumomab, cetuximab,bevacizumab, gemtuzumab, alemtuzumab, and BL22, and any fragment,derivative, conjugate, variant, radioisotope-labeled complex, orbiosimilar thereof.

In any of the methods described herein, the immunotherapeutic agents(e.g., the monoclonal antibodies) or combinations thereof may beadministered before, concurrently, or after administration of thecompounds of formulas I, II, III, IV, V, VI, VII, and/or VIII, and/orthe chemotherapeutic agents described herein.

PD-1 and PD-L1 Inhibitors

In some embodiments, the methods and/or compositions described hereininclude a combination of an HDAC inhibitor (e.g., a selective HDACinhibitor) and one or more immunotherapeutic agents. In someembodiments, the immunotherapeutic agents may include one or moreprogrammed death-1 (PD-1) and programmed death ligand 1 (PD-L1)inhibitors.

In some embodiments, the PD-1 or PD-L1 inhibitor (e.g., an ant-PD-1antibody) for use in combination with HDAC inhibitor (e.g., a selectiveHDAC inhibitor) is selected from the group consisting of nivolumab,pembrolizumab, pidilizumab, durvalumab, atezolizumab, avelumab, and anyfragment, derivative, conjugate, variant, radioisotope-labeled complex,or biosimilar thereof.

In an embodiment, an anti-PD-1 antibody comprises nivolumab (also knownas OPDIVO and commercially available from Bristol-Myers Squibb Co.), orbiosimilars, antigen-binding fragments, conjugates, or variants thereof.Nivolumab is referred to as 5C4 in International Patent Publication No.WO 2006/121168. Nivolumab is assigned Chemical Abstracts Service (CAS)registry number 946414-94-4 and is also known as BMS-936558, MDX-1106 orONO-4538. Nivolumab is a fully human IgG4 antibody blocking the PD-1receptor. The clinical safety and efficacy of nivolumab in various formsof cancer has been described in Wang et al., Cancer Immunol Res. 2014,2, 846-56; Page et al., Ann. Rev. Med., 2014, 65, 185-202; and Weber etal., J. Clin. Oncology, 2013, 31, 4311-4318. The nivolumab monoclonalantibody includes a heavy chain given by SEQ ID NO:1 and a light chaingiven by SEQ ID NO:2. Nivolumab has intra-heavy chain disulfide linkagesat 22-96, 140-196, 254-314, 360-418, 22′-96″, 140″-196″, 254″-314″, and360″-418″; intra-light chain disulfide linkages at 23′-88′, 134′-194′,23′″-88′″, and 134′″-194′″; inter-heavy-light chain disulfide linkagesat 127-214′, 127″-214′″, inter-heavy-heavy chain disulfide linkages at219-219″ and 222-222″; and N-glycosylation sites (H CH₂ 84.4) at 290,290″.

In an embodiment, the anti-PD-1 antibody is an anti-PD-1 biosimilarmonoclonal antibody approved by drug regulatory authorities withreference to nivolumab. In an embodiment, the biosimilar comprises ananti-PD-1 antibody comprising an amino acid sequence which has at least97% sequence identity, e.g., 97%, 98%, 99% or 100% sequence identity, tothe amino acid sequence of a reference medicinal product or referencebiological product and which comprises one or more post-translationalmodifications as compared to the reference medicinal product orreference biological product, wherein the reference medicinal product orreference biological product is nivolumab. In some embodiments, the oneor more post-translational modifications are selected from one or moreof: glycosylation, oxidation, deamidation, and truncation. In someembodiments, the biosimilar is an anti-PD-1 antibody authorized orsubmitted for authorization, wherein the anti-PD-1 antibody is providedin a formulation which differs from the formulations of a referencemedicinal product or or reference biological product, wherein thereference medicinal product or reference biological product isnivolumab. The anti-PD-1 antibody may be authorized by a drug regulatoryauthority such as the U.S. FDA and/or the European Union's EMA. In someembodiments, the biosimilar is provided as a composition which furthercomprises one or more excipients, wherein the one or more excipients arethe same or different to the excipients comprised in a referencemedicinal product or reference biological product, wherein the referencemedicinal product or reference biological product is nivolumab. In someembodiments, the biosimilar is provided as a composition which furthercomprises one or more excipients, wherein the one or more excipients arethe same or different to the excipients comprised in a referencemedicinal product or reference biological product, wherein the referencemedicinal product or reference biological product is nivolumab.

In an embodiment, the anti-PD-1 antibody is an antibody disclosed and/orprepared according to U.S. Pat. No. 8,008,449 or U.S. Patent ApplicationPublication Nos. 2009/0217401 A1 or 2013/0133091 A1, the disclosures ofwhich are specifically incorporated by reference herein. For example, inan embodiment, the monoclonal antibody includes 5C4 (referred to hereinas nivolumab), 17D8, 2D3, 4H1, 4A11, 7D3, and 5F4, described in U.S.Pat. No. 8,008,449, the disclosures of which are hereby incorporated byreference. The PD-1 antibodies 17D8, 2D3, 4H1, 5C4, and 4A11, are alldirected against human PD-1, bind specifically to PD-1 and do not bindto other members of the CD28 family. The sequences and CDR regions forthese antibodies are provided in U.S. Pat. No. 8,008,449, in particularin FIG. 1 through FIG. 12; the disclosures of which are incorporated byreference herein.

In another embodiment, the anti-PD-1 antibody comprises pembrolizumab(also known as KEYTRUDA), which is commercially available from Merck, orantigen-binding fragments, conjugates, or variants thereof Pembrolizumabis assigned CAS registry number 1374853-91-4 and is also known aslambrolizumab, MK-3475, and SCH-900475. The structure, properties, uses,and preparation of pembrolizumab are described in International PatentPublication No. WO 2008/156712 A1, U.S. Pat. No. 8,354,509 and U.S.Patent Application Publication Nos. US 2010/0266617 A1, US 2013/0108651A1, and US 2013/0109843 A2, the disclosures of which are incorporatedherein by reference. Pembrolizumab has an immunoglobulin G4, anti-(humanprotein PDCD1 (programmed cell death 1)) (human-Mus musculus monoclonalheavy chain), disulfide with human-Mus musculus monoclonal light chain,dimer structure. The structure of pembrolizumab may also be described asimmunoglobulin G4, anti-(human programmed cell death 1); humanized mousemonoclonal [228-L-proline (H10-S>P)]γ4 heavy chain (134-218′)-disulfidewith humanized mouse monoclonal κ light chain dimer(226-226″:229-229″)-bisdisulfide. The clinical safety and efficacy ofpembrolizumab in various forms of cancer is described in Fuerst,Oncology Times, 2014, 36, 35-36; Robert et al., Lancet, 2014, 384,1109-17; and Thomas et al., Exp. Opin. Biol. Ther., 2014, 14, 1061-1064.In an embodiment, the pembrolizumab monoclonal antibody includes a heavychain given by SEQ ID NO:11 and a light chain given by SEQ ID NO:12, andincludes the following disulfide bridges: 22-96, 22″-96″, 23′-92′,23′″-92′″, 134-218′, 134″-218′″, 138′-198′, 138′″-198′″, 147-203,147″-203″, 226-226″, 229-229″, 261-321, 261″-321″, 367-425, and367″-425″, and the following glycosylation sites (N): Asn-297 andAsn-297″. Pembrolizumab is an IgG4/kappa isotype with a stabilizingS228P mutation in the Fc region; insertion of this mutation in the IgG4hinge region prevents the formation of half molecules typically observedfor IgG4 antibodies. Pembrolizumab is heterogeneously glycosylated atAsn297 within the Fc domain of each heavy chain, yielding a molecularweight of approximately 149 kDa for the intact antibody. The dominantglycoform of pembrolizumab is the fucosylated agalacto diantennaryglycan form (GOF).

In an embodiment, the anti-PD-1 antibody is an anti-PD-1 biosimilarmonoclonal antibody approved by drug regulatory authorities withreference to pembrolizumab. In an embodiment, the biosimilar comprisesan anti-PD-1 antibody comprising an amino acid sequence which has atleast 97% sequence identity, e.g., 97%, 98%, 99% or 100% sequenceidentity, to the amino acid sequence of a reference medicinal product orreference biological product and which comprises one or morepost-translational modifications as compared to the reference medicinalproduct or reference biological product, wherein the reference medicinalproduct or reference biological product is pembrolizumab. In someembodiments, the one or more post-translational modifications areselected from one or more of: glycosylation, oxidation, deamidation, andtruncation. In some embodiments, the biosimilar is an anti-PD-1 antibodyauthorized or submitted for authorization, wherein the anti-PD-1antibody is provided in a formulation which differs from theformulations of a reference medicinal product or or reference biologicalproduct, wherein the reference medicinal product or reference biologicalproduct is pembrolizumab. The anti-PD-1 antibody may be authorized by adrug regulatory authority such as the U.S. FDA and/or the EuropeanUnion's EMA. In some embodiments, the biosimilar is provided as acomposition which further comprises one or more excipients, wherein theone or more excipients are the same or different to the excipientscomprised in a reference medicinal product or reference biologicalproduct, wherein the reference medicinal product or reference biologicalproduct is pembrolizumab. In some embodiments, the biosimilar isprovided as a composition which further comprises one or moreexcipients, wherein the one or more excipients are the same or differentto the excipients comprised in a reference medicinal product orreference biological product, wherein the reference medicinal product orreference biological product is pembrolizumab.

In an embodiment, the anti-PD-1 antibody is an antibody disclosed inU.S. Pat. No. 8,354,509 or U.S. Patent Application Publication Nos.2010/0266617 A1, 2013/0108651 A1, 2013/0109843 A2, the disclosures ofwhich are specifically incorporated by reference herein.

In an embodiment, the anti-PD-1 antibody is pidilizumab, which is alsoknown as CT-011 (CureTech Ltd.), and which is disclosed in U.S. Pat. No.8,686,119 B2, the disclosures of which are specifically incorporated byreference herein. The efficacy of pidilizumab in the treatment ofcancers, such as hematological malignancies, is described in Berger, etal., Clin. Cancer Res. 2008, 14, 3044-51. The pidilizumab monoclonalantibody includes a heavy chain given by SEQ ID NO:21 and a light chaingiven by SEQ ID NO:22. Pidilizumab has intra-heavy chain disulfidelinkages at 22-96, 144-200, 261-321, 367-425, 22″-96″, 144″-200″,261″-321″, and 367″-425″; intra-light chain disulfide linkages at23′-87′, 133′-193′, 23′″-87′″, and 133′″-193′″; inter-heavy-light chaindisulfide linkages at 220-213′ and 220″-213′″, inter-heavy-heavy chaindisulfide linkages at 226-226″ 229-229″; and N-glycosylation sites (HCH₂ 84.4) at 297, 297″.

In an embodiment, the anti-PD-1 antibody is an anti-PD-1 biosimilarmonoclonal antibody approved by drug regulatory authorities withreference to pidilizumab. In an embodiment, the biosimilar comprises ananti-PD-1 antibody comprising an amino acid sequence which has at least97% sequence identity, e.g., 97%, 98%, 99% or 100% sequence identity, tothe amino acid sequence of a reference medicinal product or referencebiological product and which comprises one or more post-translationalmodifications as compared to the reference medicinal product orreference biological product, wherein the reference medicinal product orreference biological product is pidilizumab. In some embodiments, theone or more post-translational modifications are selected from one ormore of: glycosylation, oxidation, deamidation, and truncation. In someembodiments, the biosimilar is an anti-PD-1 antibody authorized orsubmitted for authorization, wherein the anti-PD-1 antibody is providedin a formulation which differs from the formulations of a referencemedicinal product or or reference biological product, wherein thereference medicinal product or reference biological product ispidilizumab. The anti-PD-1 antibody may be authorized by a drugregulatory authority such as the U.S. FDA and/or the European Union'sEMA. In some embodiments, the biosimilar is provided as a compositionwhich further comprises one or more excipients, wherein the one or moreexcipients are the same or different to the excipients comprised in areference medicinal product or reference biological product, wherein thereference medicinal product or reference biological product ispidilizumab. In some embodiments, the biosimilar is provided as acomposition which further comprises one or more excipients, wherein theone or more excipients are the same or different to the excipientscomprised in a reference medicinal product or reference biologicalproduct, wherein the reference medicinal product or reference biologicalproduct is pidilizumab.

In an embodiment, anti-PD-1 antibodies and other PD-1 inhibitors includethose described in U.S. Pat. Nos. 8,287,856, 8,580,247, and 8,168,757and U.S. Patent Application Publication Nos. 2009/0028857 A1,2010/0285013 A1, 2013/0022600 A1, and 2011/0008369 A1, the teachings ofwhich are hereby incorporated by reference. In another embodiment,antibodies that compete with any of these antibodies for binding to PD-1are also included. In another embodiment, the anti-PD-1 antibody is anantibody disclosed in U.S. Pat. No. 8,735,553 B1, the disclosures ofwhich are incorporated herein by reference.

In an embodiment, the anti-PD-1 antibody is a commercially-availablemonoclonal antibody, such as anti-m-PD-1 clones J43 (Cat #BE0033-2) andRMP1-14 (Cat #BE0146) (Bio X Cell, Inc.). A number ofcommercially-available anti-PD-1 antibodies are known to one of ordinaryskill in the art.

In an embodiment, the PD-1 inhibitor may be a small molecule or apeptide, or a peptide derivative, such as those described in U.S. Pat.Nos. 8,907,053; 9,096,642; and 9,044,442 and U.S. Patent ApplicationPublication No. 2015/0087581; 1,2,4 oxadiazole compounds and derivativessuch as those described in U.S. Patent Application Publication No.2015/0073024; cyclic peptidomimetic compounds and derivatives such asthose described in U.S. Patent Application Publication No. 2015/0073042;cyclic compounds and derivatives such as those described in U.S. PatentApplication Publication No. 2015/0125491; 1,3,4 oxadiazole and 1,3,4thiadiazole compounds and derivatives such as those described inInternational Patent Application Publication No. WO 2015/033301;peptide-based compounds and derivatives such as those described inInternational Patent Application Publication Nos. WO 2015/036927 and WO2015/04490, or a macrocyclic peptide-based compounds and derivativessuch as those described in U.S. Patent Application Publication No.2014/0294898; the disclosures of each of which are hereby incorporatedby reference in their entireties.

The anti-PD-1 antibody sequences discussed and referenced in some of theforegoing embodiments are summarized in Table 1.

TABLE 1 Anti-PD-1 antibody amino acid sequences. IdentifierSequence (One-Letter Amino Acid Symbols) SEQ ID NO: 1QVQLVESGGG VVQPGRSLRL DCKASGITFS NSGMHWVRQA PGKGLEWVAV IWYDGSKRYY  60nivolumabADSVKGRFTI SRDNSKNTLF LQMNSLRAED TAVYYCATND DYWGQGTLVT VSSASTKGPS 120heavy chainVFPLAPCSRS TSESTAALGC LVKDYFPEPV TVSWNSGALT SGVHTFPAVL QSSGLYSLSS 180VVTVPSSSLG TKTYTCNVDH KPSNTKVDKR VESKYGPPCP PCPAPEFLGG PSVFLFPPKP 240KDTLMISRTP EVTCVVVDVS QEDPEVQFNW YVDGVEVHNA KTKPREEQFN STYRVVSVLT 300VLHQDWLNGK EYKCKVSNKG LPSSIEKTIS KAKGQPREPQ VYTLPPSQEE MTKNQVSLTC 360LVKGFYPSDI AVEWESNGQP ENNYKTTPPV LDSDGSFFLY SRLTVDKSRW QEGNVFSCSV 420MHEALHNHYT QKSLSLSLGK                                             440SEQ ID NO: 2EIVLTQSPAT LSLSPGERAT LSCRASQSVS SYLAWYQQKP GQAPRLLIYD ASNRATGIPA  60nivolumabRFSGSGSGTD FTLTISSLEP EDFAVYYCQQ SSNWPRTFGQ GTKVEIKRTV AAPSVFIFPP 120light chainSDEQLKSGTA SVVCLLNNFY PREAKVQWKV DNALQSGNSQ ESVTEQDSKD STYSLSSTLT 180LSKADYEKHK VYACEVTHQG LSSPVTKSFH RGEC                             214SEQ ID NO: 3QVQLVESGGG VVQPGRSLRL DCKASGITFS NSGMHWVRQA PGKGLEWVAV IWYDGSKRYY  60nivolumabADSVKGRFTI SRDNSKNTLF LQMNSLRAED TAVYYCATND DYWGQGTLVT VSS        113variable heavy chain SEQ ID NO: 4EIVLTQSPAT LSLSPGERAT LSCRASQSVS SYLAWYQQKP GQAPRLLIYD ASNRATGIPA  60nivolumabRFSGSGSGTD FTLTISSLEP EDFAVYYCQQ SSNWPRTFGQ GTKVEIK               107variable light chain SEQ ID NO: 5NSGMH                                                               5nivolumab heavy chain CDR1 SEQ ID NO: 6VIWYDGSKRY YADSVKG                                                 17nivolumab heavy chain CDR2 SEQ ID NO: 7NDDY                                                                4nivolumab heavy chain CDR3 SEQ ID NO: 8RASQSVSSYL A                                                       11nivolumab light chain CDR1 SEQ ID NO: 9DASNRAT                                                             7nivolumab light chain CDR2 SEQ ID NO: 10QQSSNWPRT                                                           9nivolumab light chain CDR3 SEQ ID NO: 11QVQLVQSGVE VKKPGASVKV SCKASGYTFT NYYMYWVRQA PGQGLEWMGG INPSNCCTNF  60pembrolizumabNEKFKNRVTL TTDSSTTTAY MELKSLQFDD TAVYYCARRD YRFDMGFDYW GQGTTVTVSS 120heavy chainASTKGPSVFP LAPCSRSTSE STAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSS 180GLYSLSSVVT VPSSSLGTKT YTCNVDHKPS NTKVDKRVES KYGPPCPPCP APEFLGGPSV 240FLFPPKPKDT LMISRTPEVT CVVVDVSQED PEVQFNWYVD GVEVHNAKTK PREEQFNSTY 300RVVSVLTVLH QDWLNGKEYK CKVSNKGLPS SIEKTISKAK GQPREPQVYT LPPSQEEMTK 360NQVSLTCLVK GFYPSDIAVE WESNGQPENN YKTTPPVLDS DGSFFLYSRL TVDKSRWQEG 420NVFSCSVMHE ALHNHYTQKS LSLSLGK                                     447SEQ ID NO: 12EIVLTQSPAT LSLSPGERAT LSCRASKGVS TSGYSYLHWY QQKPGQAPRL LIYLASYLES  60pembrolizumabGVPARFSGSG SGTDFTLTIS SLEPEDFAVY YCQHSRDLPL TFGGGTKVEI KRTVAAPSVF 120light chainIFPPSDEQLK SGTASVVCLL NNFYPREAKV QWKVDNALQS GNSQESVTEQ DSKDSTYSLS 180STLTLSKADY EKHKVYACEV THQGLSSPVT KSFNRGEC                         218SEQ ID NO: 13QVQLVQSGVE VKKPGASVKV SCKASGYTFT NYYMYWVRQA PGQGLEWMGG INPSNGGTNF  60pembrolizumabNEKFKNRVTL TTDSSTTTAY MELKSLQFDD TAVYYCARRD YRFDMGFDYW GQGTTVTVSS 120variable heavy chain SEQ ID NO: 14EIVLTQSPAT LSLSPGERAT LSCRASKGVS TSGYSYLHWY QQKPGQAPRL LIYLASYLES  60pembrolizumabGVPARFSGSG SGTDFTLTIS SLEPEDFAVY YCQHSRDLPL TFGGGTKVEI K          111variable light chain SEQ ID NO: 15NYYMY                                                               5pemb rolinumab heavy chain CDR1 SEQ ID NO: 16GINPSNGGTN FNEKFK                                                  16pembrolizumab heavy chain CDR2 SEQ ID NO: 17RDYRFDKGFD Y                                                       11pembrolizumab heavy chain CDR3 SEQ ID NO: 18RASKGVSTSG YSYLH                                                   15pembrolizumab light chain CDR1 SEQ ID NO: 19LASYLES                                                             7pembrolizumab light chain CDR2 SEQ ID NO: 20QHSRDLPLT                                                           9pembrolizumab light chain CDR3 SEQ ID NO: 21QVQLVQSGSE LKKPGASVKI SCKASGYTFT NYGKNWVRQA PGQGLQWMGW INTDSGESTY  60pidilizumabAEEFKGRFVF SLDTSVNTAY LQITSLTAED TGMYFCVRVG YDALDYWGQG TLVTVSSAST 120heavy chainKCPSVFPLAP SSKSTSGGTA ALGCLVKDYF PEPVTVSWNS GALTSGVHTF PAVLQSSGLY 180SLSSVVTVPS SSLGTQTYIC NVNHKPSNTK VDKRVEPKSC CKTHTCPPCP APELLGGPSV 240FLFPPKPKDT LMISRTPEVT CVVVDVSHED PEVKFNWYVD GVEVHNAKTK PREEQYNSTY 300RVVSVLTVLH QDWLNGKEYK CKVSNKALPA PIEKTISKAK GQPREPQVYT LPPSREEMTK 360NQVSLTCLVK GFYPSCIAVE WESNGQPENN YKTTPPVLDS CGSFFLYSKL TVDKSRWQQG 420NVFSCSVMHE ALHNHYTQKS LSLSPGK                                     447SEQ ID NO: 22EIVLTQSPSS LSASVGDRVT ITCSARSSVS YKHWFQQKPG KAPKLWIYRT SNLASGVPSR  60pidilizumabFSGSGSGTSY CLTINSLQPE DFATYYCQQR SSFPLTFGGG TKLEIKRTVA APSVFIFPPS 120light chainDEQLKSGTAS VVCLLNNFYP RFAKVQWKVD NALQSGNSQE SVTEQDSKDS TYSLSSTLTL 180SKADYEKHKV YACEVTHQGL SSPVTKSFNR GEC                              213SEQ ID NO: 23QVQLVQSGSE LKKPGASVKI SCKASGYTFT NYGKNWVRQA PGQGLQWMGW INTDSGESTY  60pidilizumabAEEFKGRFVF SLDTSVNTAY LQITSLTAED TGMYFCVRVG YDALDYWGQG TLVTVSS    117variable heavy chain SEQ ID NO: 24EIVLTQSPSS LSASVGDRVT ITCSARSSVS YMHWFQQKPG KAPKLWIYRT SNLASGVPSR  60pidilizumabFSGSGSGTSY CLTINSLQPE DFATYYCQQR SSFPLTFGGG TKLEIK                106variable light chain

The PD-L1 inhibitor may be any PD-L1 inhibitor or blocker known in theart. In particular, it is one of the PD-L1 inhibitors or blockersdescribed in more detail in the following paragraphs. The terms“inhibitor” and “blocker” are used interchangeably herein in referenceto PD-L1 inhibitors. For avoidance of doubt, references herein to aPD-L1 inhibitor that is an antibody may refer to a compound or fragment,derivative, conjugate, variant, radioisotope-labeled complex, orbiosimilar thereof. For avoidance of doubt, references herein to a PD-L1inhibitor may refer to a compound or a pharmaceutically acceptable salt,ester, solvate, hydrate, cocrystal, or prodrug thereof.

In an embodiment, the anti-PD-L1 antibody is durvalumab, also known asMEDI4736 (which is commercially available from Medimmune, LLC), orantigen-binding fragments, conjugates, or variants thereof. In anembodiment, the anti-PD-L1 antibody is an antibody disclosed in U.S.Pat. No. 8,779,108 or U.S. Patent Application Publication No.2013/0034559, the disclosures of which are specifically incorporated byreference herein. The clinical efficacy of durvalumab (MEDI4736, SEQ IDNO:25 and SEQ ID NO:26) has been described in: Page et al., Ann. Rev.Med., 2014, 65, 185-202; Brahmer et al., J. Clin. Oncol. 2014, 32, 5s(supplement, abstract 8021); and McDermott et al., Cancer TreatmentRev., 2014, 40, 1056-64. The durvalumab monoclonal antibody includesdisulfide linkages at 22-96, 22″-96″, 23′-89′, 23′″-89′″, 135′-195′,135′″-195′″, 148-204, 148″-204″, 215′-224, 215′″-224″, 230-230″,233-233″, 265-325, 265″-325″, 371-429, and 371″-429′; andN-glycosylation sites at Asn-301 and Asn-301″.

In an embodiment, the anti-PD-L1 antibody is an anti-PD-L1 biosimilarmonoclonal antibody approved by drug regulatory authorities withreference to durvalumab. In an embodiment, the biosimilar comprises ananti-PD-L1 antibody comprising an amino acid sequence which has at least97% sequence identity, e.g., 97%, 98%, 99% or 100% sequence identity, tothe amino acid sequence of a reference medicinal product or referencebiological product and which comprises one or more post-translationalmodifications as compared to the reference medicinal product orreference biological product, wherein the reference medicinal product orreference biological product is durvalumab. In some embodiments, the oneor more post-translational modifications are selected from one or moreof: glycosylation, oxidation, deamidation, and truncation. In someembodiments, the biosimilar is an anti-PD-L1 antibody authorized orsubmitted for authorization, wherein the anti-PD-L1 antibody is providedin a formulation which differs from the formulations of a referencemedicinal product or or reference biological product, wherein thereference medicinal product or reference biological product isdurvalumab. In some embodiments, the biosimilar is provided as acomposition which further comprises one or more excipients, wherein theone or more excipients are the same or different to the excipientscomprised in a reference medicinal product or reference biologicalproduct, wherein the reference medicinal product or reference biologicalproduct is durvalumab. In some embodiments, the biosimilar is providedas a composition which further comprises one or more excipients, whereinthe one or more excipients are the same or different to the excipientscomprised in a reference medicinal product or reference biologicalproduct, wherein the reference medicinal product or reference biologicalproduct is durvalumab.

In an embodiment, anti-PD-L1 antibodies and other PD-L1 inhibitorsinclude those described in U.S. Pat. No. 8,779,108 and U.S. PatentApplication Publication No. 2013/0034559A1, the disclosures of which arehereby incorporated by reference. In another embodiment, antibodies thatcompete with any of these antibodies for binding to PD-L1 are alsoincluded.

In an embodiment, the anti-PD-L1 antibody is atezolizumab, also known asMPDL3280A or RG7446 (commercially available from Genentech, Inc.), orantigen-binding fragments, conjugates, or variants thereof. In anembodiment, the anti-PD-L1 antibody is an antibody disclosed in U.S.Pat. No. 8,217,149, the disclosure of which is specifically incorporatedby reference herein. In an embodiment, the anti-PD-L1 antibody is anantibody disclosed in U.S. Patent Application Publication Nos.2010/0203056 A1, 2013/0045200 A1, 2013/0045201 A1, 2013/0045202 A1, or2014/0065135 A1, the disclosures of which are specifically incorporatedby reference herein. The atezolizumab monoclonal antibody includes aheavy chain given by SEQ ID NO:35 and a light chain given by SEQ IDNO:36. Atezolizumab has intra-heavy chain disulfide linkages (C23-C104)at 22-96, 145-201, 262-322, 368-426, 22″-96″, 145″-201″, 262″-322″, and368″-426″; intra-light chain disulfide linkages (C23-C104) at 23′-88′,134′-194′, 23′″-88′″, and 134′″-194′″; intra-heavy-light chain disulfidelinkages (h 5-CL 126) at 221-214′ and 221″-214′″; intra-heavy-heavychain disulfide linkages (h 11, h 14) at 227-227″ and 230-230″; andN-glycosylation sites (H CH₂ N84.4>A) at 298 and 298′.

In an embodiment, the anti-PD-L1 antibody is an anti-PD-L1 biosimilarmonoclonal antibody approved by drug regulatory authorities withreference to atezolizumab. In an embodiment, the biosimilar comprises ananti-PD-L1 antibody comprising an amino acid sequence which has at least97% sequence identity, e.g., 97%, 98%, 99% or 100% sequence identity, tothe amino acid sequence of a reference medicinal product or referencebiological product and which comprises one or more post-translationalmodifications as compared to the reference medicinal product orreference biological product, wherein the reference medicinal product orreference biological product is atezolizumab. In some embodiments, theone or more post-translational modifications are selected from one ormore of: glycosylation, oxidation, deamidation, and truncation. In someembodiments, the biosimilar is an anti-PD-L1 antibody authorized orsubmitted for authorization, wherein the anti-PD-L1 antibody is providedin a formulation which differs from the formulations of a referencemedicinal product or or reference biological product, wherein thereference medicinal product or reference biological product isatezolizumab. The anti-PD-L1 antibody may be authorized by a drugregulatory authority such as the U.S. FDA and/or the European Union'sEMA. In some embodiments, the biosimilar is provided as a compositionwhich further comprises one or more excipients, wherein the one or moreexcipients are the same or different to the excipients comprised in areference medicinal product or reference biological product, wherein thereference medicinal product or reference biological product isatezolizumab. In some embodiments, the biosimilar is provided as acomposition which further comprises one or more excipients, wherein theone or more excipients are the same or different to the excipientscomprised in a reference medicinal product or reference biologicalproduct, wherein the reference medicinal product or reference biologicalproduct is atezolizumab.

In an embodiment, the anti-PD-L1 antibody is avelumab, also known asMSB0010718C (commercially available from Merck KGaA/EMD Serono), orantigen-binding fragments, conjugates, or variants thereof. In anembodiment, the anti-PD-L1 antibody is an antibody disclosed in U.S.Patent Application Publication No. US 2014/0341917 A1, the disclosure ofwhich is specifically incorporated by reference herein. The avelumabmonoclonal antibody includes a heavy chain given by SEQ ID NO:45 and alight chain given by SEQ ID NO:46. Avelumab has intra-heavy chaindisulfide linkages (C23-C104) at 22-96, 147-203, 264-324, 370-428,22″-96″, 147″-203″, 264″-324″, and 370″-428″; intra-light chaindisulfide linkages (C23-C104) at 22′-90′, 138′-197′, 22′″-90′″, and138′″-197′″; intra-heavy-light chain disulfide linkages (h 5-CL 126) at223-215′ and 223″-215′″; intra-heavy-heavy chain disulfide linkages (h11, h 14) at 229-229″ and 232-232″; N-glycosylation sites (H CH₂ N84.4)at 300, 300″; fucosylated complex bi-antennary CHO-type glycans; and HCHS K2 C-terminal lysine clipping at 450 and 450′.

In an embodiment, the anti-PD-L1 antibody is an anti-PD-L1 biosimilarmonoclonal antibody approved by drug regulatory authorities withreference to avelumab. In an embodiment, the biosimilar comprises ananti-PD-L1 antibody comprising an amino acid sequence which has at least97% sequence identity, e.g., 97%, 98%, 99% or 100% sequence identity, tothe amino acid sequence of a reference medicinal product or referencebiological product and which comprises one or more post-translationalmodifications as compared to the reference medicinal product orreference biological product, wherein the reference medicinal product orreference biological product is avelumab. In some embodiments, the oneor more post-translational modifications are selected from one or moreof: glycosylation, oxidation, deamidation, and truncation. In someembodiments, the biosimilar is an anti-PD-L1 antibody authorized orsubmitted for authorization, wherein the anti-PD-L1 antibody is providedin a formulation which differs from the formulations of a referencemedicinal product or or reference biological product, wherein thereference medicinal product or reference biological product is avelumab.The anti-PD-L1 antibody may be authorized by a drug regulatory authoritysuch as the U.S. FDA and/or the European Union's EMA. In someembodiments, the biosimilar is provided as a composition which furthercomprises one or more excipients, wherein the one or more excipients arethe same or different to the excipients comprised in a referencemedicinal product or reference biological product, wherein the referencemedicinal product or reference biological product is avelumab. In someembodiments, the biosimilar is provided as a composition which furthercomprises one or more excipients, wherein the one or more excipients arethe same or different to the excipients comprised in a referencemedicinal product or reference biological product, wherein the referencemedicinal product or reference biological product is avelumab.

In an embodiment, anti-PD-L1 antibodies and other PD-L1 inhibitorsinclude those described in U.S. Patent Application Publication No.2014/0341917 A1, the disclosure of which is hereby incorporated byreference. In another embodiment, antibodies that compete with any ofthese antibodies for binding to PD-L1 are also included.

In an embodiment, the anti-PD-L1 antibody is MDX-1105, also known asBMS-935559, which is disclosed in U.S. Pat. No. 7,943,743 B2, thedisclosures of which are specifically incorporated by reference herein.In an embodiment, the anti-PD-L1 antibody is selected from theanti-PD-L1 antibodies disclosed in U.S. Pat. No. 7,943,743 B2, which arespecifically incorporated by reference herein.

In an embodiment, the anti-PD-L1 antibody is a commercially-availablemonoclonal antibody, such as INVIVOMAB anti-m-PD-L1 clone 10F.9G2(Catalog #BE0101, Bio X Cell, Inc.). In an embodiment, the anti-PD-L1antibody is a commercially-available monoclonal antibody, such asAFFYMETRIX EBIOSCIENCE (MIH1). A number of commercially-availableanti-PD-L1 antibodies are known to one of ordinary skill in the art.

The anti-PD-L1 antibody sequences referenced in some of the foregoingembodiments are summarized in Table 2.

TABLE 2 Anti-PD-L1 antibody amino acid sequences. IdentifierSequence (One-Letter Amino Acid Symbols) SEQ ID NO: 25EVQLVESGGG LVQPGGSLRL SCAASGFTFS RYWMSWVRQA PGKGLEWVAN IKQDGSEKYY  60durvalumabVDSVKGRFTI SRDNAKNSLY LQMNSLRAED TAVYYCAREG GWFGELAFDY WGQGTLVTVS 120(MEDI4736)SASTKGPSVF PLAPSSKSTS GGTAALGCLV KDYFPEPVTV SWNSGALTSG VHTFPAVLQS 180heavy chainSGLYSLSSVV TVPSSSLGTQ TYICNVNHKP SNTKVDKRVE PKSCDKTHTC PPCPAPEFEG 240GPSVFLFPPK PKDTLMISRT PEVTCVVVDV SHEDPEVKFN WYVDGVEVHN AKTKPREEQY 300NSTYRVVSVL TVLHQDWLNG KEYKCKVSNK ALPASIEKTI SKAKGQPREP QVYTLPPSRE 360EMTKNQVSLT CLVKGFYPSD IAVEWESNGQ PENNYKTTPP VLDSDGSFFL YSKLTVDKSR 420WQQGNVFSCS VMHEALHNHY TQKSLSLSPG K                                451SEQ ID NO: 26EVQLVESGGG LVQPGGSLRL SCAASGFTFS RYWMSWVRQA PGKGLEWVAN EIVLTQSPGT  60durvalumabLSLSPGERAT LSCRASQRVS SSYLAWYQQK PGQAPRLLIY DASSRATGIP DRFSGSGSGT 120(MEDI4736)DFTLTISRLE PEDFAVYYCQ QYGSLPWTFG QGTKVEIKRT VAAPSVFIFP PSDEQLKSGT 180light chainASVVCLLNNF YPREAKVQWK VDNALQSGNS QESVTEQDSK DSTYSLSSTL TLSKADYEKH 240KVYACEVTHQGLSSPVTKSF NRGEC                                                  265SEQ ID NO: 27EVQLVESGGG LVQPGGSLRL SCAASGFTFS RYWMSWVRQA PGKGLEWVAN IKQDGSEKYY  60durvalumabVDSVKGRFTI SRDNAKNSLY LQMNSLRAED TAVYYCAREG GWFGELAFDY WGQGTLVTVS 120variableS                                                                 121heavy chain SEQ ID NO: 28EIVLTQSPGT LSLSPGERAT LSCRASQRVS SSYLAWYQQK PGQAPRLLIY DASSRATGIP  60durvalumabDRFSGSGSGT DFTLTISRLE PEDFAVYYCQ QYGSLPWTFG QGTKVEIK              108variable light chain SEQ ID NO: 29RYWMS                                                               5durvalumab heavy chain CDR1 SEQ ID NO: 30NIKQDGSEKY YVDSVKG                                                 17durvalumab heavy chain CDR2 SEQ ID NO: 31EGGWFGELAF DY                                                      12durvalumab heavy chain CDR3 SEQ ID NO: 32RASQRVSSSY LA                                                      12durvalumab light chain CDR1 SEQ ID NO: 33DASSRAT                                                             7durvalumab light chain CDR2 SEQ ID NO: 34QQYGSLPWT                                                           9durvalumab light chain CDR3 SEQ ID NO: 35EVQLVESGGG LVQPGGSLRL SCAASGFTFS DSWIHWVRQA PGKGLEWVAW ISPYGGSTYY  60atezolizumabADSVKGRFTI SADTSKNTAY LQMNSLRAED TAVYYCARRH WPGGFDYWGQ GTLVTVSSAS 120(MPDL3280A)TKGPSVFPLA PSSKSTSGGT AALGCLVKDY FPEPVTVSWN SGALTSGVHT FPAVLQSSGL 180heavy chainYSLSSVVTVP SSSLGTQTYI CNVNHKPSNT KVDKKVEPKS CDKTHTCPPC PAPELLGGPS 240VFLFPPKPKD TLMISRTPEV TCVVVDVSHE DPEVKFNWYV DGVEVHNAKT KPREEQYAST 300YRVVSVLTVL HQDWLNGKEY KCKVSNKALP APIEKTISKA KGQPREPQVY TLPPSREEMT 360KNQVSLTCLV KGFYPSDIAV EWESNGQPEN NYKTTPPVLD SDGSFFLYSK LTVDKSRWQQ 420GNVFSCSVMH EALHNHYTQK SLSLSPGK                                    448SEQ ID NO: 36DIQMTQSPSS LSASVGDRVT ITCRASQDVS TAVAWYQQKP GKAPKLLIYS ASFLYSGVPS  60atezolizumabRFSGSGSGTD FTLTISSLQP EDFATYYCQQ YLYHPATFGQ GTKVEIKRTV AAPSVFIFPP 120(MPDL3280A)SDEQLKSGTA SVVCLLNNFY PREAKVQWKV DNALQSGNSQ ESVTEQDSKD STYSLSSTLT 180light chainLSKADYEKHK VYACEVTHQG LSSPVTKSFN RGEC                             214SEQ ID NO: 37EVQLVESGGG LVQPGGSLRL SCAASGFTFS DSWIHWVRQA PGKGLEWVAW ISPYGGSTYY  60atezolizumabADSVKGRFTI SADTSKNTAY LQMNSLRAED TAVYYCARRH WPGGFDYWGQ GTLVTVSA   118variable heavy chain SEQ ID NO: 38DIQMTQSPSS LSASVGDRVT ITCRASQDVS TAVAWYQQKP GKAPKLLIYS ASFLYSGVPS  60atezolizumabRFSGSGSGTD FTLTISSLQP EDFATYYCQQ YLYHPATFGQ GTKVEIKR              108variable light chain SEQ ID NO: 39GFTFSXSWIH                                                         10atezolizumab heavy chain CDR1 SEQ ID NO: 40AWIXPYGGSX YYADSVKG                                                18atezolizumab heavy chain CDR2 SEQ ID NO: 41RHWPGGFDY                                                           9atezolizumab heavy chain CDR3 SEQ ID NO: 42RASQXXXTXX A                                                       11atezolizumab light chain CDR1 SEQ ID NO: 43SASXLXS                                                             7atezolizumab light chain CDR2 SEQ ID NO: 44QQXXXXPXT                                                           9atezolizumab light chain CDR3 SEQ ID NO: 45EVQLLESGGG LVQPGGSLRL SCAASGFTFS SYIMMWVRQA PGKGLEWVSS IYPSGGITFY  60avelumabADTVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARIK LGTVTTVDYW GQGTLVTVSS 120(MSB0010718C)ASTKGPSVFP LAPSSKSTSG GTAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSS 180heavy chainGLYSLSSVVT VPSSSLGTQT YICNVNHKPS NTKVDKKVEP KSCDKTHTCP PCPAPELLGG 240PSVFLFPPKP KDTLMISRTP EVTCVVVDVS HEDPEVKFNW YVDGVEVHNA KTKPREEQYN 300STYRVVSVLT VLHQDWLNGK EYKCKVSNKA LPAPIEKTIS KAKGQPREPQ VYTLPPSRDE 360LTKNQVSLTC LVKGFYPSDI AVEWESNGQP ENNYKTTPPV LDSDGSFFLY SKLTVDKSRW 420QQGNVFSCSV MHEALHNHYT QKSLSLSPGK                                  450SEQ ID NO: 46QSALTQPASV SGSPGQSITI SCTGTSSDVG GYNYVSWYQQ HPGKAPKLMI YDVSNRPSGV  60avelumabSNRFSGSKSG NTASLTISGL QAEDEADYYC SSYTSSSTRV FGTGTKVTVL GQPKANPTVT 120(MSB0010718C)LFPPSSEELQ ANKATLVCLI SDFYPGAVTV AWKADGSPVK AGVETTKPSK QSNNKYAASS 180light chainYLSLTPEQWK SHRSYSCQVT HEGSTVEKTV APTECS                           216SEQ ID NO: 47EVQLLESGGG LVQPGGSLRL SCAASGFTFS SYIMMWVRQA PGKGLEWVSS IYPSGGITFY  60avelumabADTVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARIK LGTVTTVDYW GQGTLVTVSS 120variable heavy chain SEQ ID NO: 48QSALTQPASV SGSPGQSITI SCTGTSSDVG GYNYVSWYQQ HPGKAPKLMI YDVSNRPSGV  60avelumabSNRFSGSKSG NTASLTISGL QAEDEADYYC SSYTSSSTRV FGTGTKVTVL            110variable light chain SEQ ID NO: 49SYIMM                                                               5avelumab heavy chain CDR1 SEQ ID NO: 50SIYPSGGITF YADTVKG                                                 17avelumab heavy chain CDR2 SEQ ID NO: 51IKLGTVTTVD Y                                                       11avelumab heavy chain CDR3 SEQ ID NO: 52TGTSSDVGGY NYVS                                                    14avelumab light chain CDR1 SEQ ID NO: 53DVSNRPS                                                             7avelumab light chain CDR2 SEQ ID NO: 54SSYTSSSTRV                                                         10avelumab light chain CDR3

The preparation, properties, and uses of suitable PD-1 and PD-L1inhibitors are described in, e.g., U.S. Pat. No. 8,008,449 or U.S.Patent Application Publication Nos. 2009/0217401 A1 or 2013/0133091 A1;U.S. Pat. No. 8,354,509 and U.S. Patent Application Publication Nos.2010/0266617 A1, 2013/0108651 A1, and 2013/0109843 A2; U.S. Pat. Nos.8,287,856, 8,580,247, and 8,168,757 and U.S. Patent ApplicationPublication Nos. US 2009/0028857 A1, US 2010/0285013 A1, US 2013/0022600A1, and US 2011/0008369 A1; U.S. Pat. No. 8,779,108 or U.S. PatentApplication Publication No. 2013/0034559 A1; U.S. Pat. No. 8,217,149 andU.S. Patent Application Publication Nos. 2010/0203056 A1, 2013/0045200A1, 2013/0045201 A1, 2013/0045202 A1, or 2014/0065135 A1; and U.S.Patent Application Publication No. 2014/0341917 A1, the disclosures ofeach of which are incorporated by reference herein.

In an embodiment, a PD-1 or PD-L1 inhibitor selected from the groupconsisting of nivolimumab, pembrolizumab, pidilizumab, durvalumab,atezolizumab, avelumab, and/or Fab fragments, antigen-binding fragments,derivatives, conjugates, variants, and radioisotope-labeled complexesthereof, is administered to a subject by infusing a dose selected fromthe group consisting of about 10 mg, about 20 mg, about 25 mg, about 50mg, about 75 mg, 100 mg, about 200 mg, about 300 mg, about 400 mg, about500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about1000 mg, about 1100 mg, about 1200 mg, about 1300 mg, about 1400 mg,about 1500 mg, about 1600 mg, about 1700 mg, about 1800 mg, about 1900mg, and about 2000 mg. In an embodiment, the PD-1 or PD-L1 inhibitor isadministered weekly. In an embodiment, the PD-1 or PD-L1 inhibitor isadministered every two weeks. In an embodiment, the PD-1 or PD-L1inhibitor is administered every three weeks. In an embodiment, the PD-1or PD-L1 inhibitor is administered monthly. In an embodiment, the PD-1or PD-L1 inhibitor is administered at a lower initial dose, which isescalated when administered at subsequent intervals administeredmonthly.

Other Immunotherapeutic Agents

In some embodiments, the methods and/or compositions described hereininclude a combination of an HDAC inhibitor (e.g., a selective HDACinhibitor) and one or more immunotherapeutic agents. In someembodiments, the immunotherapeutic agents may include one or more ofrituximab (Rituxan®; SEQ ID NO.55 (heavy chain) and SEQ ID NO.56 (lightchain)), trastuzumab (Herceptin®; SEQ ID NO.57 (heavy chain) and SEQ IDNO.58 (light chain)), ibritumomab (Zevalin®; SEQ ID NO.59 (heavy chain)and SEQ ID NO.60 (light chain)), tositumomab (Bexxar®; SEQ ID NO.61(heavy chain) and SEQ ID NO.62 (light chain)), cetuximab (C-225,Erbitux®; SEQ ID NO.63 (heavy chain) and SEQ ID NO.64 (light chain)),bevacizumab (Avastin®; SEQ ID NO.65 (heavy chain) and SEQ ID NO.66(light chain)), gemtuzumab (Mylotarg®; SEQ ID NO.67 (heavy chain) andSEQ ID NO.68 (light chain)), alemtuzumab (Campath®; SEQ ID NO.69 (heavychain) and SEQ ID NO.70 (light chain)), and BL22 (RFB4 (dsFv)-PE38 orCAT-3888, and as described in Mansfield, et al., Blood (1997) 90:2020-2026 and Kreitman, et al., Clin Cancer Res. (2011) 17: 6398-6405).

In an embodiment, the immunotherapeutic agent comprises one or more ofrituximab, trastuzumab, ibritumomab, tositumomab, cetuximab,bevacizumab, gemtuzumab, alemtuzumab, and BL22, or a biosimilar,antigen-binding fragment, derivative, conjugate, variant, orradioisotope-labeled complex thereof.

In an embodiment, the forementioned immunotherapeutic agents may beprovided in a pharmaceutical composition or method, as described herein,as a biosimilar monoclonal antibody approved by drug regulatoryauthorities with reference to rituximab, trastuzumab, ibritumomab,tositumomab, cetuximab, bevacizumab, gemtuzumab, alemtuzumab, or BL22.In an embodiment, the biosimilar comprises a monoclonal antibodycomprising an amino acid sequence which has at least 97% sequenceidentity, e.g., 97%, 98%, 99% or 100% sequence identity, to the aminoacid sequence of a reference medicinal product or reference biologicalproduct and which comprises one or more post-translational modificationsas compared to the reference medicinal product or reference biologicalproduct, wherein the reference medicinal product or reference biologicalproduct is rituximab, trastuzumab, ibritumomab, tositumomab, cetuximab,bevacizumab, gemtuzumab, alemtuzumab, or BL22. In some embodiments, theone or more post-translational modifications are selected from one ormore of: glycosylation, oxidation, deamidation, and truncation. In someembodiments, the biosimilar is an monoclonal antibody authorized orsubmitted for authorization, wherein the monoclonal antibody is providedin a formulation which differs from the formulations of a referencemedicinal product or or reference biological product, wherein thereference medicinal product or reference biological product isrituximab, trastuzumab, ibritumomab, tositumomab, cetuximab,bevacizumab, gemtuzumab, alemtuzumab, or BL22. The monoclonal antibodymay be authorized by a drug regulatory authority such as the U.S. FDAand/or the European Union's EMA. In some embodiments, the biosimilar isprovided as a composition which further comprises one or moreexcipients, wherein the one or more excipients are the same or differentto the excipients comprised in a reference medicinal product orreference biological product, wherein the reference medicinal product orreference biological product is rituximab, trastuzumab, ibritumomab,tositumomab, cetuximab, bevacizumab, gemtuzumab, alemtuzumab, or BL22.In some embodiments, the biosimilar is provided as a composition whichfurther comprises one or more excipients, wherein the one or moreexcipients are the same or different to the excipients comprised in areference medicinal product or reference biological product, wherein thereference medicinal product or reference biological product isrituximab, trastuzumab, ibritumomab, tositumomab, cetuximab,bevacizumab, gemtuzumab, alemtuzumab, or BL22.

In an embodiment, a monoclonal antibody selected from the groupconsisting of rituximab, trastuzumab, ibritumomab, tositumomab,cetuximab, bevacizumab, gemtuzumab, alemtuzumab, and BL22, and/orbiosimilars, antigen-binding fragments, derivatives, conjugates,variants, and radioisotope-labeled complexes thereof, is administered toa subject by infusing a dose selected from the group consisting of about10 mg, about 20 mg, about 25 mg, about 50 mg, about 75 mg, 100 mg, about200 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about700 mg, about 800 mg, about 900 mg, about 1000 mg, about 1100 mg, about1200 mg, about 1300 mg, about 1400 mg, about 1500 mg, about 1600 mg,about 1700 mg, about 1800 mg, about 1900 mg, and about 2000 mg. In anembodiment, the monoclonal antibody is administered weekly. In anembodiment, the monoclonal antibody is administered every two weeks. Inan embodiment, the monoclonal antibody is administered every threeweeks. In an embodiment, the monoclonal antibody is administeredmonthly. In an embodiment, the monoclonal antibody is administered at alower initial dose, which is escalated when administered at subsequentintervals administered monthly

Several of the other immunotherapeutic agent sequences referenced in theforegoing sections are summarized in Table 3.

TABLE 3 Other immunotherapeutic agent antibody sequences. SequenceIdentifier and Description Sequence (One-Letter Amino Acid Symbols)SEQ ID NO: 55QVQLQQPGAE LVKPGASVKM SCKASGYTFT SYNMHWVKQT PGRGLEWIGA IYPGNGDTSY  60rituximabNQKFKGKATL TADKSSSTAY MQLSSLTSED SAVYYCARST YYGGDWYFNV WGAGTTVTVS 120heavy chainAASTKGPSVF PLAPSSKSTS GGTAALGCLV KDYFPEPVTV SWNSGALTSG VHTFPAVLQS 180SGLYSLSSVV TVPSSSLGTQ TYICNVNHKP SNTKVDKKVE PKSCDKTHTC PPCPAPELLG 240GPSVFLFPPK PKDTLMISRT PEVTCVVVDV SHEDPEVKFN WYVDGVEVHN AKTKPREEQY 300NSTYRVVSVL TVLHQDWLNG KEYKCKVSNK ALPAPIEKTI SKAKGQPREP QVYTLPPSRD 360ELTKNQVSLT CLVKGFYPSD IAVEWESNGQ PENNYKTTPP VLDSDGSFFL YSKLTVDKSR 420WQQGNVFSCS VMHEALHNHY TQKSLSLSPG K                                451SEQ ID NO: 56QIVLSQSPAI LSASPGEKVT MTCRASSSVS YIHWFQQKPG SSPKPWIYAT SNLASGVPVR  60rituximabFSGSGSGTSY SLTISRVEAE DAATYYCQQW TSNPPTFGGG TKLEIKRTVA APSVFIFPPS 120light chainDEQLKSGTAS VVCLLNNFYP REAKVQWKVD NALQSGNSQE SVTEQDSKDS TYSLSSTLTL 180SKADYEKHKV YACEVTHQGL SSPVTKSFNR GEC                              213SEQ ID NO: 57EVQLVESGGG LVQPGGSLRL SCAASGFNIK DTYIHWVRQA PGKGLEWVAR IYPTNGYTRY  60trastuzumabADSVKGRFTI SADTSKNTAY LQMNSLRAED TAVYYCSRWG GDGFYAMDYW GQGTLVTVSS 120heavy chainASTKGPSVFP LAPSSKSTSG GTAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSS 180GLYSLSSVVT VPSSSLGTQT YICNVNHKPS NTKVDKKVEP PKSCDKTHTC PPCPAPELLG 240GPSVFLFPPK PKDTLMISRT PEVTCVVVDV SHEDPEVKFN WYVDGVEVHN AKTKPREEQY 300NSTYRVVSVL TVLHQDWLNG KEYKCKVSNK ALPAPIEKTI SKAKGQPREP QVYTLPPSRD 360ELTKNQVSLT CLVKGFYPSD IAVEWESNGQ PENNYKTTPP VLDSDGSFFL YSKLTVDKSR 420WQQGNVFSCS VMHEALHNHY TQKSLSLSPG K                                451SEQ ID NO: 58DIQMTQSPSS LSASVGDRVT ITCRASQDVN TAVAWYQQKP GKAPKLLIYS ASFLYSGVPS  60trastuzumabRFSGSRSGTD FTLTISSLQP EDFATYYCQQ HYTTPPTFGQ GTKVEIKRTV AAPSVFIFPP 120light chainSDEQLKSGTA SVVCLLNNFY PREAKVQWKV DNALQSGNSQ ESVTEQDSKD STYSLSSTLT 180LSKADYEKHK VYACEVTHQG LSSPVTKSFN RGEC                             214SEQ ID NO: 59QAYLQQSGAE LVRPGASVKM SCKASGYTFT SYNMHWVKQT PRQGLEWIGA IYPGNGDTSY  60ibritumomabNQKFKGKATL TVDKSSSTAY MQLSSLTSED SAVYFCARVV YYSNSYWYFD VWGTGTTVTV 120heavy chainSAPSVYPLAP VCGDTTGSSV TLGCLVKGYF PEPVTLTWNS GSLSSGVHTF PAVLQSDLYT 180LSSSVTVTSS TWPSQSITCN VAHPASSTKV DKKIEPRGPT IKPCPPCKCP APNLLGGPSV 240FIFPPKIKDV LMISLSPIVT CVVVDVSEDD PDVQISWFVN NVEVHTAQTQ THREDYNSTL 300RVVSALPIQH QDWMSGKEFK CKVNNKDLPA PIERTISKPK GSVRAPQVYV LPPPEEEMTK 360KQVTLTCMVT DFMPEDIYVE WTNNGKTELN YKNTEPVLDS DGSYFMYSKL RVEKKNWVER 420NSYSCSVVHE GLHNHHTTKS FSR                                         443SEQ ID NO: 60QIVLSQSPAI LSASPGEKVT MTCRASSSVS YMHWYQQKPG SSPKPWIYAP SNLASGVPAR  60ibritumomabFSGSGSGTSY SLTISRVEAE DAATYYCQQW SFNPPTFGAG TKLELKRADA APTVFIFPPS 120light chainDEQLKSGTAS VVCLLNNFYP REAKVQWKVD NALQSGNSQE SVTEQDSKDS TYSLSSTLTL 180SKADYEKHKV YACEVTHQGL SSPVTKSFN                                   209SEQ ID NO: 61QAYLQQSGAE LVRPGASVKM SCKASGYTFT SYNMHWVKQT PRQGLEWIGA IYPGNGDTSY  60tositumomabNQKFKGKATL TVDKSSSTAY MQLSSLTSED SAVYFCARVV YYSNSYWYFD VWGTGTTVTV 120heavy chainSGPSVFPLAP SSKSTSGGTA ALGCLVKDYF PEPVTVSWNS GALTSGVHTF PAVLQSSGLY 180SLSSVVTVPS SSLGTQTYIC NVNHKPSNTK VDKKAEPKSC DKTHTCPPCP APELLGGPSV 240FLFPPKPKDT LMISRTPEVT CVVVDVSHED PEVKFNWYVD GVEVHNAKTK PREEQYNSTY 300RVVSVLTVLH QDWLNGKEYK CKVSNKALPA PIEKTISKAK GQPREPQVYT LPPSRDELTK 360NQVSLTCLVK GFYPSDIAVE WESNGQPENN YKTTPPVLDS DGSFFLYSKL TVDKSRWQQG 420NVFSCSVMHE ALHNHYTQKS LSLSPGK                                     447SEQ ID NO: 62QIVLSQSPAI LSASPGEKVT MTCRASSSVS YMHWYQQKPG SSPKPWIYAP SNLASGVPAR  60tositumomabFSGSGSGTSY SLTISRVEAE DAATYYCQQW SFNPPTFGAG TKLELKRTVA APSVFIFPPS 120light chainDEQLKSGTAS VVCLLNNFYP REAKVQWKVD NALQSGNSQE SVTEQDSKDS TYSLSSTLTL 180SKADYEKHKV YACEVTHQGL SSPVTKSFNR                                  210SEQ ID NO: 63QVQLKQSGPG LVQPSQSLSI TCTVSGFSLT NYGVHWVRQS PGKGLEWLGV IWSGGNTDYN  60cetuximabTPFTSRLSIN KDNSKSQVFF KMNSLQSNDT AIYYCARALT YYDYEFAYWG QGTLVTVSAA 120heavy chainSTKGPSVFPL APSSKSTSGG TAALGCLVKD YFPEPVTVSW NSGALTSGVH TFPAVLQSSG 180LYSLSSVVTV PSSSLGTQTY ICNVNHKPSN TKVDKRVEPK SPKSCDKTHT CPPCPAPELL 240GGPSVFLFPP KPKDTLMISR TPEVTCVVVD VSHEDPEVKF NWYVDGVEVH NAKTKPREEQ 300YNSTYRVVSV LTVLHQDWLN GKEYKCKVSN KALPAPIEKT ISKAKGQPRE PQVYTLPPSR 360DELTKNQVSL TCLVKGFYPS DIAVEWESNG QPENNYKTTP PVLDSDGSFF LYSKLTVDKS 420RWQQGNVFSC SVMHEALHNH YTQKSLSLSP GK                               452SEQ ID NO: 64DILLTQSPVI LSVSPGERVS FSCRASQSIG TNIHWYQQRT NGSPRLLIKY ASESISGIPS  60cetuximabRFSGSGSGTD FTLSINSVES EDIADYYCQQ NNNWPTTFGA GTKLELKRTV AAPSVFIFPP 120light chainSDEQLKSGTA SVVCLLNNFY PREAKVQWKV DNALQSGNSQ ESVTEQDSKD STYSLSSTLT 180LSKADYEKHK VYACEVTHQG LSSPVTKSFN RGA                              213SEQ ID NO: 65EVQLVESGGG LVQPGGSLRL SCAASGYTFT NYGMNWVRQA PGKGLEWVGW INTYTGEPTY  60bevacizumabAADFKRRFTF SLDTSKSTAY LQMNSLRAED TAVYYCAKYP HYYGSSHWYF DVWGQGTLVT 120heavy chainVSSASTKGPS VFPLAPSSKS TSGGTAALGC LVKDYFPEPV TVSWNSGALT SGVHTFPAVL 180QSSGLYSLSS VVTVPSSSLG TQTYICNVNH KPSNTKVDKK VEPKSCDKTH TCPPCPAPEL 240LGGPSVFLFP PKPKDTLMIS RTPEVTCVVV DVSHEDPEVK FNWYVDGVEV HNAKTKPREE 300QYNSTYRVVS VLTVLHQDWL NGKEYKCKVS NKALPAPIEK TISKAKGQPR EPQVYTLPPS 360REEMTKNQVS LTCLVKGFYP SDIAVEWESN GQPENNYKTT PPVLDSDGSF FLYSKLTVDK 420SRWQQGNVFS CSVMHEALHN HYTQKSLSLS PGK                              453SEQ ID NO: 66DIQMTQSPSS LSASVGDRVT ITCSASQDIS NYLNWYQQKP GKAPKVLIYF TSSLHSGVPS  60bevacizumabRFSGSGSGTD FTLTISSLQP EDFATYYCQQ YSTVPWTFGQ GTKVEIKRTV AAPSVFIFPP 120Light chainSDEQLKSGTA SVVCLLNNFY PREAKVQWKV DNALQSGNSQ ESVTEQDSKD STYSLSSTLT 180LSKADYEKHK VYACEVTHQG LSSPVTKSFN RGEC                             214SEQ ID NO: 67QVQLQQSGAE LAKPGASVKM SCKASGYTFT SYRMHWVKQR PGQGLEWIGY INPSTGYTEY  60gemtuzumabNQKFKDKATL TADKSSSTAY MQLSSLTFED SAVYYCARGG GVFDYWGQGT TLTVSS     116heavy chain SEQ ID NO: 68QIVLTQSPAI MSASPGEKVT ITCSASSSIS YMHWFQQKPG TSPKLWIYTT SNLASGVPAR  60gemtuzumabFSGSGSGTSY SLTISRMEAE DAATYYCHQR STYPLTFGSG TKLELK                106Light chain SEQ ID NO: 69QVQLQESGPG LVRPSQTLSL TCTVSGFTFT DFYMNWVRQP PGRGLEWIGF IRDKAKGYTT  60alemtuzumabEYNPSVKGRV TMLVDTSKNQ FSLRLSSVTA ADTAVYYCAR EGHTAAPFDY WGQGSLVTVS 120heavy chainSASTKGPSVF PLAPSSKSTS GGTAALGCLV KDYFPEPVTV SWNSGALTSG VHTFPAVLQS 180SGLYSLSSVV TVPSSSLGTQ TYICNVNHKP SNTKVDKKVE PKSCDKTHTC PPCPAPELLG 240GPSVFLFPPK PKDTLMISRT PEVTCVVVDV SHEDPEVKFN WYVDGVEVHN AKTKPREEQY 300NSTYRVVSVL TVLHQDWLNG KEYKCKVSNK ALPAPIEKTI SKAKGQPREP QVYTLPPSRD 360ELTKNQVSLT CLVKGFYPSD IAVEWESNGQ PENNYKTTPP VLDSDGSFFL YSKLTVDKSR 420WQQGNVFSCS VMHEALHNHY TQKSLSLSPG K                                451SEQ ID NO: 70DIQMTQSPSS LSASVGDRVT ITCKASQNID KYLNWYQQKP GKAPKLLIYN TNNLQTGVPS  60alemtuzumabRFSGSGSGTD FTFTISSLQP EDIATYYCLQ HISRPRTFGQ GTKVEIKRTV AAPSVFIFPP 120Light chainSDEQLKSGTA SVVCLLNNFY PREAKVQWKV DNALQSGNSQ ESVTEQDSKD STYSLSSTLT 180LSKADYEKHK VYACEVTHQG LSSPVTKSFN R                                211Methods of Treating Cancer

Cancer is the second leading cause of death in the United States afterheart disease. The American Cancer Society estimates that 1,665,540 newcancer cases are expected to have been diagnosed in 2014 with 585,720cancer-related deaths.

In some embodiments, the invention includes methods of treating adisease alleviated by inhibiting histone deacetylase (HDAC) protein in apatient in need thereof, wherein the treatment includes the step ofadministering a therapeutically effective amount of one or more HDACinhibitors described herein (e.g., one or more HDAC inhibitors describedby formulas I, II, III, IV, V, VI, VII, and/or VIII). In someembodiments, the disease is cancer. For example, the cancer may be oneof the cancers described in Table 4.

In some embodiments, the one or more HDAC inhibitors may include HDACinhibitors that selectively inhibit HDAC 6. For example, the HDACinhibitors used in the methods of the invention may include a compoundof formula IV, VII, and/or VIII, or a pharmaceutically acceptable salt,solvate, hydrate, cocrystal, or prodrug thereof.

In some embodiments, the methods of the invention may includeadministering a therapeutically effective amount of an additionaltherapeutic agent such as, for example, chemotherapeutic agents and/orimmunotherapeutic agents, as described herein.

In some embodiments, a patient responding to the methods of treatmentdisclosed herein may exhibit the absence of disease progression (e.g.,halting the growth and/or spread of neoplastic cells and tissues) overanother patient that does not receive the methods of treatment describedherein.

In some embodiments, the cancer treated by the methods and/orcompositions described herein may include those cancers listed in Table4.

TABLE 4 Selected cancers that may be treated by the methods of theinvention. Exemplary Solid Tumors: acoustic neuroma adenocarcinomaangiosarcoma astrocytoma basal cell carcinoma bile duct carcinomabladder carcinoma breast cancer bronchogenic carcinoma cervical cancerchordoma choriocarcinoma colon cancer colorectal cancer craniopharygiomacystadenocarcinoma embryonal carcinoma endotheliosarcoma ependymomaepithelial carcinoma esophagaelcancer Ewing's tumor fibrosarcomaglioblastomamultiforme glioma hemangioblastoma hepatoma kidney cancerleiomyosarcoma liposarcoma lung cancer lymphangioendotheliosarcomalymphangiosarcoma medullary carcinoma medulloblastoma melanomameningioma mesothelioma myxosarcoma nasal cancer neuroblastomaoligodendroglioma oral cancer osteogenic sarcoma ovarian cancerpancreatic cancer papillary adenocarcinomas papillary carcinomapinealoma prostate cancer rabdomyosarcoma renal cell carcinomaretinoblastoma sebaceous gland carcinoma seminoma skin cancer squamouscell carcinoma stomach cancer sweat gland carcinoma synovioma testicularcancer small cell lung carcinoma throat cancer uterine cancer Wilms'tumor Exemplary Blood Cancers: acute erythroleukemic leukemia acutelymphoblastic B-cell leukemia acute lymphoblastic T-cell leukemia acutelypmhoblastic leukemia acute megakaryoblastic leukemia acute monoblasticleukemia acute myeloblastic leukemia acute myelomonocytic leukemia acutenonlymphocytic leukemia acute promyelocytic leukemia acuteundifferentiated leukemia chronic lymphocytic leukemia chronicmyelocytic leukemia hairy cell leukemia multiple myeloma ExemplaryLymphomas: heavy chain disease Hodgkin's disease multiple myelomanon-Hodgkin's lymphoma polycythemia vera Waldenstrom's macroglobulinemia

In some embodiments, the cancers described herein that may be treated bythe methods of the invention may be treated with or without a furtherstep of irradiation.

In some embodiments, the cancer treated by the methods of the inventionmay include acoustic neuroma, adenocarcinoma, angiosarcoma, astrocytoma,basal cell carcinoma, bile duct carcinoma, bladder carcinoma, braincancer, breast cancer, brochogenic carcinoma, cervical cancer, chordoma,choriocarcinoma, colon cancer, colorectal cancer, craniopharygioma,cystadenocarcinoma, embryonal carcinoma, endotheliocarconima,ependymoma, epithelial carcinoma, esophageal cancer, Ewing's tumor,fibrosarcoma, gastric cancer, glioblastoma multiforme, glioma, head andneck cancer, hemangioblastoma, hepatoma, kidney cancer, leiomyosarcoma,liposarcoma, lung cancer, lymphangioendotheliosarcoma,lymphangiosarcoma, medullary carcinoma, medulloblastoma, melanoma,meningioma, mesothelioma, myxosarcoma, nasal cancer, neuroblastoma,oligodendroglioma, oral cancer, osteogenic sarcoma, ovarian cancer,pancreatic cancer, papillary adenocarcinoma, papillary carcinoma,pinealoma, prostate cancer, rabdomyosarcoma, rectal cancer, renal cellcarcinoma, retinoblastoma, sarcoma, sebacaceous gland carcinoma,seminoma, skin cancer, squamous cell carcinoma, stomach cancer, sweatgland carcinoma, synovioma, testicular cancer, small cell lungcarcinoma, throat cancer, uterine cancer, Wilm's tumor, blood cancer,acute erythroleukemic leukemia, acute lymphoblastic B-cell leukemia,acute lymphoblastic T-cell leukemia, acute lymphoblastic leukemia, acutemegakaryoblastic leukemia, acute monoblastic leukemia, acutemyeloblastic leukemia, acute myelomonocytic leukemia, acutenonlymphocytic leukemia, acute promyelocytic leukemia, acuteundifferentiated leukemia, chronic lymphocytic leukemia, chronicmyelocytic leukemia, hairy cell leukemia, multiple myeloma, heavy chaindisease, Hodgkin's disease, multiple myeloma, non-Hodgkin's lymphoma,polycythemia vera, and Waldenstrom's macroglobulinemia.

In some embodiments, the cancers treated by the methods and/orcompositions of the invention are HPV positive (+) cancers. HPV (+)cancers may include cervical cancer and head and neck cancer.

In some embodiments, treatments of cancer may include surgery,radiotherapy, and/or chemotherapy. Each treatment modality carries risksand benefits, and cancer recurrences underlie efforts to improve theoutcomes of treatment. In particular, recent advances in surgical andradiation therapy technologies, employing computational and roboticmethods, have plateaued efficacy of local-regional treatments. Moreover,targeted agents to personalize chemotherapy have altered the cancertreatment paradigm.

Radiation therapy (i.e., radiotherapy) involves the treatment of cancerand other diseases using ionizing radiation. Ionizing radiation depositsenergy that injures or destroys cells in targeted tissues by damagingtheir genetic material and subsequently interfering with a cell'sability to grow and/or replicate. Radiation exposure damages cancercells and normal cells, but the normal cells activate processes tobetter repair themselves and may continue to function properly.Radiotherapy may be used to treat solid tumors (e.g., cancers of thehead and neck, breast, prostate, rectum, uterus, lung, brain, kidney,uterus, and cervix). Radiotherapy may also be used to treat cancers suchas leukemias and lymphomas. Radiotherapies used for leukemias andlymphomas may include total body radiation therapy in protocolspreparing patients for bone marrow transplants. Radiotherapy may be moreeffective when the targeted cancer tissues are more sensitive to theeffects of radiation than surrounding normal tissues.

The radiation responses of different cancers or tumors may vary as afunction of histology, cellular doubling time, oxygenation, nutrientavailability, repair capacity, and other factors. Some cancers arereadily cured using ionizing radiation doses within normal tissuetolerances, while other types of cancer may not be very responsive toradiation. Furthermore, radiation responses of tumors with the samehistology may show considerable heterogeneity and reduce the therapeuticeffects of the therapy. Thus, a primary challenge facing radiotherapy isthe differentiation between the more radiosensitive tumors versus lessradiosensitive tumors and the surrounding healthy tissues.

Investigations into the molecular bases underlying cellular radiationresponses have provided dramatic mechanistic insight. Signaltransduction pathways have been implicated to play important roles incellular responses to ionizing radiation. Induction of gene expressionby these cascades under various conditions has been shown to result incell cycle arrest, activation of DNA repair processes, and activation ofprogrammed cell death (apoptosis). Disruption of critical signalingpathways in cancer cells may result in enhanced cytotoxic effectsfollowing radiation exposure. Certain cells may be disrupted byinterfering with the histone acetylation and deacetylation processes ofthe cells.

Therapeutic ratios may be determined by measuring the effects of drugson cancers and on normal tissues. Radiation toxicities to organs at riskmay affect normal tissues adjacent to the treated volume (such as rectumor bladder in the treatment of a pelvic tumor), or in sites receivingtransit dose (such as the pelvic bone marrow).

The methods of the invention may include irradiating a selected tissueof the patient before, during, and/or after a compound of the invention(or pharmaceutical composition containing such compound) is administeredto the patient. Regarding the application of radiation (“radiationtherapy” or “radiotherapy”) to the patient or subject more generally,such therapy may encompass any ionizing radiation known to those havingordinary skill in the art. Generally, radiation therapy, and inparticular ionizing radiation includes applying to a selected tissue,such as a selected tissue comprising cancerous and/or neoplastic cells,a dose of ionizing radiation or two or more fractions of ionizingradiation. The ionization radiation is defined as an irradiation dosewhich is determined according to the disease's characteristics at theselected tissue and therapeutic decision of a physician. The term“fractionated dose(s)” may include, for example, conventionalfractionation, hyperfractionation, hypofractionation, and acceleratedfractionation). The amount of radiation and doses thereof should besufficient to damage the highly proliferating cells' genetic material,making it impossible for the irradiated cells to continue growing anddividing.

In certain aspects, fractionated irradiation may vary from daily doses(e.g. one or more times per day) given for a period of weeks, or to onceweekly doses given for a period of weeks or months. Indeed, radiationmay be applied in dosages of about 0.1 Gy to about 100 Gy. For example,the dosage may be about 5 to 15 Gy.

In certain fractionated irradiation methods, irradiation dosing mayinclude the application of about 0.1 to about 20 Gy or from about 1 Gyto about 10 Gy or from about 1 Gy to about 3 Gy in a single session,which may be repeated several times over the course of about 1 to 10weeks, or about 2 to 5 weeks. In certain embodiments of the invention,the radiation dose may be about 30 to 60 Gy at 1 to 5 Gy fractions overa period of about 2 to 5 weeks.

In other exemplary aspects, three different fractionation schemes may beused in accordance with the invention.

In one embodiment, radiation doses from 1 Gy to 3 Gy in daily fractionsfor several weeks (e.g., about 2 to 8 weeks) to achieve cumulative dosesof about 20 Gy to 80 Gy.

In another embodiment, large fraction radiation therapy may includedoses of 4 Gy to 25 Gy. This fractionated irradiation scheme may includethe delivery of about 1 fraction to 5 fractions delivered over about 1-2weeks. This type of radiation may be referred to as stereotacticradiosurgery or stereotactic body radiation therapy.

In a further embodiment, brachytherapy may be used, which is deliveredusing low dose and rate techniques or high-dose rate techniques,typically delivering doses of about 4 Gy to 10 Gy per day with techniqueand fractionation specific to the clinical situation as would beunderstood by a person having ordinary skill in the art.

As set forth above, the compounds and/or compositions of the inventionmay be administered before, after, or together with the radiation. Onecycle of radiation therapy as well as several cycles of radiation ispossible, dependent on the reduction of tumor size or extent ofproliferation. Such sequences of radiosensitization treatments andionizing irradiation are repeated as needed to abate and, optimally,reduce or eliminate the spread of the cancer or neoplastic cells in thetissue or region of tissue that is selected for treatment. Accordingly,the total dose and the radiation regimen will depend, inter alia, on thecancer type, type of compound that results in radiosensitization,irradiated area, physical condition of the patient and many otherconsiderations appreciated by those having ordinary skill in the art.

In addition to the administration of a compound of the invention and theirradiation of the patient, the methods of the invention may include theadministration of a therapeutically effective amount of achemotherapeutic agent and/or an immunotherapeutic agent to the patient.The chemotherapeutic agent and/or immunotherapeutic agent may beprovided before, during, or after at least one of the steps ofadministering the HDAC inhibitor and irradiating a selected tissue ofthe patient. Therefore, the chemotherapeutic agent and/orimmunotherapeutic agent may be provided at various points during themethods of the invention for the treatment of disease. In certainaspects, the chemotherapeutic agent and/or immunotherapeutic agent maybe administered concurrently with or after the step of irradiating theselected tissue of the patient.

Methods of Treating Immunological Diseases

Regarding immunological diseases, the HDAC inhibitors may be used inmethods of treating diseases that are the result of over-activeimmunity. Millions of people worldwide endure debilitating immunologicaldiseases that implicate HDAC proteins and may be treated by thecompounds, compositions, and methods of the invention. For example, seeInternational Patent Application No. 2011/017448, the entirety of whichis incorporated herein by reference.

In some embodiments, the invention includes methods of treating adisease alleviated by inhibiting histone deacetylase (HDAC) protein in apatient in need thereof, wherein the treatment includes the step ofadministering a therapeutically effective amount of one or more HDACinhibitors described herein (e.g., such as a selective HDAC inhibitor,including one or more compounds of formulas I, II, III, IV, V, VI, VII,and/or VIII). In some embodiments, the disease is an immunologicaldisease, as described herein.

In some embodiments, the immunological disease may be celiac disease,diabetes mellitus type 1 (IDDM), systemic lupus erythematosus (SLE),Sjogren's syndrome, Churg-Strauss Syndrome, Hashimoto's thyroiditis,Graves' disease, idiopathic thrombocytopenic purpura, rheumatoidarthritis (RA), polymyositis, ulcerative colitis, Crohn's disease,autoimmune carditis, Wegener's granulomatosis, autoimmune hemolyticanemia, polyarteritis nodosa, psoriasis, vitiligo, epidermolysisbullosa, scleroderma, alopecia areata, epidermolysis bullosa acquisita,bullous pemphigoid, pemphigus foliaceous, and pemphigus vulgaris.

In some embodiments, the one or more HDAC inhibitors may include HDACinhibitors that selectively inhibit HDAC 6. For example, the HDACinhibitors used in the methods of the invention may include a compoundof formula VI, VII, or VIII, or a pharmaceutically acceptable salt,solvate, hydrate, cocrystal, or prodrug thereof.

In some embodiments, the methods of the invention may includeadministering a therapeutically effective amount of an additionaltherapeutic agent such as, for example, chemotherapeutic agents and/orimmunotherapeutic agents, as described herein.

Methods of Treating Neurological Diseases

Neurological diseases affect a vast number of humans of all ages. In theUnited States, over 500,000 people each year experience a stroke, makingit the third leading cause of death and the primary cause of disability.One in twenty people is afflicted with Alzheimer's disease by the age of65, and almost 40 percent of the population have the disease by age 80.More than 600,000 people suffer from Parkinson's disease and over200,000 from multiple sclerosis. Every year, greater than 10,000 peopledie from amyotrophic lateral sclerosis (ALS). The impact of neurologicaldisease is not only devastating for the patients, but also for theirfamilies.

Although considerable effort has been invested in the design ofeffective therapies, neurological diseases continue to threaten theworldwide population and lessen their quality of life. The HDACinhibitors may be used in compositions or methods for treating suchneurological disorders that implicate HDAC proteins.

In some embodiments, the invention includes methods of treating adisease alleviated by inhibiting histone deacetylase (HDAC) protein in apatient in need thereof, wherein the treatment includes the step ofadministering a therapeutically effective amount of one or more HDACinhibitors described herein (e.g., such as a selective HDAC inhibitor,including one or more compounds of formulas I, II, III, IV, V, VI, VII,and/or VIII). In some embodiments, the disease is a neurologicaldisease, as described herein.

In some embodiments, the neurological disease may be selected from thegroup consisting of Huntington's disease, spinal muscular atrophy (SMA),Parkinson's disease, Alzheimer's, Multiple Sclerosis, and AmyotrophicLateral Sclerosis (ALS). In certain aspects, the neurological diseasemay be Alzheimer's disease or multiple sclerosis.

In some embodiments, the one or more HDAC inhibitors may include HDACinhibitors that selectively inhibit HDAC6. For example, the HDACinhibitors used in the methods of the invention may include a compoundof formula VI, VII, or VIII, or a pharmaceutically acceptable salt,solvate, hydrate, cocrystal, or prodrug thereof.

In some embodiments, the methods of the invention may includeadministering a therapeutically effective amount of an additionaltherapeutic agent such as, for example, chemotherapeutic agents and/orimmunotherapeutic agents, as described herein.

Methods of Treating Inflammatory Diseases

Compounds having a hydroxamic acid moiety are known to possess usefulbiological activities in addition to being useful for treating cancer,immunological diseases, and neurological diseases. For example, manypeptidyl compounds possessing a hydroxamic acid moiety are known toinhibit matrix metalloproteinases (MMPs), which are a family of zincendopeptidases. The MMPs play a key role in both physiological andpathological tissue degradation. Therefore, peptidyl compounds that havethe ability to inhibit the action of MMPs show utility for the treatmentor prophylaxis of conditions involving tissue breakdown andinflammation.

In some embodiments, the invention includes methods of treating adisease alleviated by inhibiting histone deacetylase (HDAC) protein in apatient in need thereof, wherein the treatment includes the step ofadministering a therapeutically effective amount of one or more HDACinhibitors described herein (e.g., such as a selective HDAC inhibitor,including one or more compounds of formulas I, II, III, IV, V, VI, VII,and/or VIII). In some embodiments, the disease is an inflammatorydisease, as described herein.

In some embodiments, the inflammatory diseases may include, for example,psoriatic arthritis; inflammatory bowel diseases such as Crohn's diseaseand ulcerative colitis; spondyloarthropathies; scleroderma; psoriasis(including T-cell mediated psoriasis) and inflammatory dermatoses suchan dermatitis, eczema, atopic dermatitis, allergic contact dermatitis,urticaria; vasculitis (e.g., necrotizing, cutaneous, andhypersensitivity vasculitis); eosinphilic myositis, eosinophilicfasciitis; acute pancreatitis; inflammatory bowel disease; or aninflammatory condition resulting from strain, sprain, cartilage damage,trauma such as burn, orthopedic surgery, infection or other diseaseprocesses. See, for example, U.S. Pat. No. 7,507,858.

In some embodiments, the one or more HDAC inhibitors may include HDACinhibitors that selectively inhibit HDAC6. For example, the HDACinhibitors used in the methods of the invention may include a compoundof formula VI, VII, or VIII, or a pharmaceutically acceptable salt,solvate, hydrate, cocrystal, or prodrug thereof.

In some embodiments, the methods of the invention may includeadministering a therapeutically effective amount of an additionaltherapeutic agent such as, for example, chemotherapeutic agents and/orimmunotherapeutic agents, as described herein.

Pharmaceutical Compositions

In an embodiment, an active pharmaceutical ingredient or combination ofactive pharmaceutical ingredients, such as any of the foregoing HDACinhibitors (e.g., such as a selective HDAC inhibitor, including one ormore compounds of formulas I, II, III, IV, V, VI, VII, and/or VIII), isprovided as a pharmaceutically acceptable composition.

In some embodiments, the concentration of each of the activepharmaceutical ingredients provided in the pharmaceutical compositionsof the invention, such as any of the foregoing HDAC inhibitors, is lessthan, for example, 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%,18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%,2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%,0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%,0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%,0.0005%, 0.0004%, 0.0003%, 0.0002% or 0.0001% w/w, w/v or v/v of thepharmaceutical composition.

In some embodiments, the concentration of each of the activepharmaceutical ingredients provided in the pharmaceutical compositionsof the invention, such as any of the foregoing HDAC inhibitors, isgreater than 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19.75%, 19.50%,19.25% 19%, 18.75%, 18.50%, 18.25% 18%, 17.75%, 17.50%, 17.25% 17%,16.75%, 16.50%, 16.25% 16%, 15.75%, 15.50%, 15.25% 15%, 14.75%, 14.50%,14.25% 14%, 13.75%, 13.50%, 13.25% 13%, 12.75%, 12.50%, 12.25% 12%,11.75%, 11.50%, 11.25% 11%, 10.75%, 10.50%, 10.25% 10%, 9.75%, 9.50%,9.25% 9%, 8.75%, 8.50%, 8.25% 8%, 7.75%, 7.50%, 7.25% 7%, 6.75%, 6.50%,6.25% 6%, 5.75%, 5.50%, 5.25% 5%, 4.75%, 4.50%, 4.25%, 4%, 3.75%, 3.50%,3.25%, 3%, 2.75%, 2.50%, 2.25%, 2%, 1.75%, 1.50%, 125%, 1%, 0.5%, 0.4%,0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%,0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%,0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%,0.0003%, 0.0002% or 0.0001% w/w, w/v, or v/v of the pharmaceuticalcomposition.

In some embodiments, the concentration of each of the activepharmaceutical ingredients provided in the pharmaceutical compositionsof the invention, such as any of the foregoing HDAC inhibitors, is inthe range from about 0.0001% to about 50%, about 0.001% to about 40%,about 0.01% to about 30%, about 0.02% to about 29%, about 0.03% to about28%, about 0.04% to about 27%, about 0.05% to about 26%, about 0.06% toabout 25%, about 0.07% to about 24%, about 0.08% to about 23%, about0.09% to about 22%, about 0.1% to about 21%, about 0.2% to about 20%,about 0.3% to about 19%, about 0.4% to about 18%, about 0.5% to about17%, about 0.6% to about 16%, about 0.7% to about 15%, about 0.8% toabout 14%, about 0.9% to about 12% or about 1% to about 10% w/w, w/v orv/v of the pharmaceutical composition.

In some embodiments, the concentration of each of the activepharmaceutical ingredients provided in the pharmaceutical compositionsof the invention, such as any of the foregoing HDAC inhibitors, is inthe range from about 0.001% to about 10%, about 0.01% to about 5%, about0.02% to about 4.5%, about 0.03% to about 4%, about 0.04% to about 3.5%,about 0.05% to about 3%, about 0.06% to about 2.5%, about 0.07% to about2%, about 0.08% to about 1.5%, about 0.09% to about 1%, about 0.1% toabout 0.9% w/w, w/v or v/v of the pharmaceutical composition.

In some embodiments, the amount of each of the active pharmaceuticalingredients provided in the pharmaceutical compositions of theinvention, such as any of the foregoing HDAC inhibitors, is equal to orless than 10 g, 9.5 g, 9.0 g, 8.5 g, 8.0 g, 7.5 g, 7.0 g, 6.5 g, 6.0 g,5.5 g, 5.0 g, 4.5 g, 4.0 g, 3.5 g, 3.0 g, 2.5 g, 2.0 g, 1.5 g, 1.0 g,0.95 g, 0.9 g, 0.85 g, 0.8 g, 0.75 g, 0.7 g, 0.65 g, 0.6 g, 0.55 g, 0.5g, 0.45 g, 0.4 g, 0.35 g, 0.3 g, 0.25 g, 0.2 g, 0.15 g, 0.1 g, 0.09 g,0.08 g, 0.07 g, 0.06 g, 0.05 g, 0.04 g, 0.03 g, 0.02 g, 0.01 g, 0.009 g,0.008 g, 0.007 g, 0.006 g, 0.005 g, 0.004 g, 0.003 g, 0.002 g, 0.001 g,0.0009 g, 0.0008 g, 0.0007 g, 0.0006 g, 0.0005 g, 0.0004 g, 0.0003 g,0.0002 g, or 0.0001 g.

In some embodiments, the amount of each of the active pharmaceuticalingredients provided in the pharmaceutical compositions of theinvention, such as any of the foregoing HDAC inhibitors, is more than0.0001 g, 0.0002 g, 0.0003 g, 0.0004 g, 0.0005 g, 0.0006 g, 0.0007 g,0.0008 g, 0.0009 g, 0.001 g, 0.0015 g, 0.002 g, 0.0025 g, 0.003 g,0.0035 g, 0.004 g, 0.0045 g, 0.005 g, 0.0055 g, 0.006 g, 0.0065 g, 0.007g, 0.0075 g, 0.008 g, 0.0085 g, 0.009 g, 0.0095 g, 0.01 g, 0.015 g, 0.02g, 0.025 g, 0.03 g, 0.035 g, 0.04 g, 0.045 g, 0.05 g, 0.055 g, 0.06 g,0.065 g, 0.07 g, 0.075 g, 0.08 g, 0.085 g, 0.09 g, 0.095 g, 0.1 g, 0.15g, 0.2 g, 0.25 g, 0.3 g, 0.35 g, 0.4 g, 0.45 g, 0.5 g, 0.55 g, 0.6 g,0.65 g, 0.7 g, 0.75 g, 0.8 g, 0.85 g, 0.9 g, 0.95 g, 1 g, 1.5 g, 2 g,2.5 g, 3 g, 3.5 g, 4 g, 4.5 g, 5 g, 5.5 g, 6 g, 6.5 g, 7 g, 7.5 g, 8 g,8.5 g, 9 g, 9.5 g, or 10 g.

Each of the active pharmaceutical ingredients according to the inventionis effective over a wide dosage range. For example, in the treatment ofadult humans, dosages independently range from 0.01 to 1000 mg, from 0.5to 100 mg, from 1 to 50 mg per day, and from 5 to 40 mg per day areexamples of dosages that may be used. The exact dosage will depend uponthe route of administration, the form in which the compound isadministered, the gender and age of the subject to be treated, the bodyweight of the subject to be treated, and the preference and experienceof the attending physician. The clinically-established dosages of theforegoing HDAC inhibitors may also be used if appropriate.

In an embodiment, the molar ratio of two active pharmaceuticalingredients in the pharmaceutical compositions is in the range from 10:1to 1:10, preferably from 2.5:1 to 1:2.5, and more preferably about 1:1.In an embodiment, the weight ratio of the molar ratio of two activepharmaceutical ingredients in the pharmaceutical compositions isselected from the group consisting of 20:1, 19:1, 18:1, 17:1, 16:1,15:1, 14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1,2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12,1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, and 1:20. In an embodiment,the weight ratio of the molar ratio of two active pharmaceuticalingredients in the pharmaceutical compositions is selected from thegroup consisting of 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1, 13:1,12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3,1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16,1:17, 1:18, 1:19, and 1:20.

Of course, as those skilled in the art will appreciate, the dosageactually administered will depend upon the condition being treated, theage, health and weight of the recipient, the type of concurrenttreatment, if any, and the frequency of treatment. Moreover, theeffective dosage amount may be determined by one skilled in the art onthe basis of routine empirical activity testing to measure thebioactivity of the compound(s) in a bioassay, and thus establish theappropriate dosage to be administered.

Furthermore, the described methods of treatment may normally includemedical follow-up to determine the therapeutic or prophylactic effectbrought about in the subject undergoing treatment with the compound(s)and/or composition(s) described herein.

Described below are non-limiting pharmaceutical compositions and methodsfor preparing the same.

Pharmaceutical Compositions for Oral Administration

In an embodiment, the invention provides a pharmaceutical compositionfor oral administration containing the active pharmaceutical ingredientor combination of active pharmaceutical ingredients, such as the HDACinhibitors described herein (e.g., such as a selective HDAC inhibitor,including one or more compounds of formulas I, II, III, IV, V, VI, VII,and/or VIII), and a pharmaceutical excipient suitable for oraladministration.

In some embodiments, the invention provides a solid pharmaceuticalcomposition for oral administration containing: (i) an effective amountof an active pharmaceutical ingredient or combination of activepharmaceutical ingredients, and (ii) a pharmaceutical excipient suitablefor oral administration. In selected embodiments, the compositionfurther contains (iii) an effective amount of a third activepharmaceutical ingredient and optionally (iv) an effective amount of afourth active pharmaceutical ingredient.

In some embodiments, the pharmaceutical composition may be a liquidpharmaceutical composition suitable for oral consumption. Pharmaceuticalcompositions of the invention suitable for oral administration can bepresented as discrete dosage forms, such as capsules, sachets, ortablets, or liquids or aerosol sprays each containing a predeterminedamount of an active ingredient as a powder or in granules, a solution,or a suspension in an aqueous or non-aqueous liquid, an oil-in-wateremulsion, a water-in-oil liquid emulsion, powders for reconstitution,powders for oral consumptions, bottles (including powders or liquids ina bottle), orally dissolving films, lozenges, pastes, tubes, gums, andpacks. Such dosage forms can be prepared by any of the methods ofpharmacy, but all methods include the step of bringing the activeingredient(s) into association with the carrier, which constitutes oneor more necessary ingredients. In general, the compositions are preparedby uniformly and intimately admixing the active ingredient(s) withliquid carriers or finely divided solid carriers or both, and then, ifnecessary, shaping the product into the desired presentation. Forexample, a tablet can be prepared by compression or molding, optionallywith one or more accessory ingredients. Compressed tablets can beprepared by compressing in a suitable machine the active ingredient in afree-flowing form such as powder or granules, optionally mixed with anexcipient such as, but not limited to, a binder, a lubricant, an inertdiluent, and/or a surface active or dispersing agent. Molded tablets canbe made by molding in a suitable machine a mixture of the powderedcompound moistened with an inert liquid diluent.

The invention further encompasses anhydrous pharmaceutical compositionsand dosage forms since water can facilitate the degradation of somecompounds. For example, water may be added (e.g., 5%) in thepharmaceutical arts as a means of simulating long-term storage in orderto determine characteristics such as shelf-life or the stability offormulations over time. Anhydrous pharmaceutical compositions and dosageforms of the invention can be prepared using anhydrous or low moisturecontaining ingredients and low moisture or low humidity conditions.Pharmaceutical compositions and dosage forms of the invention whichcontain lactose can be made anhydrous if substantial contact withmoisture and/or humidity during manufacturing, packaging, and/or storageis expected. An anhydrous pharmaceutical composition may be prepared andstored such that its anhydrous nature is maintained. Accordingly,anhydrous compositions may be packaged using materials known to preventexposure to water such that they can be included in suitable formularykits. Examples of suitable packaging include, but are not limited to,hermetically sealed foils, plastic or the like, unit dose containers,blister packs, and strip packs.

Each of the active pharmaceutical ingredients can be combined in anintimate admixture with a pharmaceutical carrier according toconventional pharmaceutical compounding techniques. The carrier can takea wide variety of forms depending on the form of preparation desired foradministration. In preparing the compositions for an oral dosage form,any of the usual pharmaceutical media can be employed as carriers, suchas, for example, water, glycols, oils, alcohols, flavoring agents,preservatives, coloring agents, and the like in the case of oral liquidpreparations (such as suspensions, solutions, and elixirs) or aerosols;or carriers such as starches, sugars, micro-crystalline cellulose,diluents, granulating agents, lubricants, binders, and disintegratingagents can be used in the case of oral solid preparations, in someembodiments without employing the use of lactose. For example, suitablecarriers include powders, capsules, and tablets, with the solid oralpreparations. If desired, tablets can be coated by standard aqueous ornonaqueous techniques.

Binders suitable for use in pharmaceutical compositions and dosage formsinclude, but are not limited to, corn starch, potato starch, or otherstarches, gelatin, natural and synthetic gums such as acacia, sodiumalginate, alginic acid, other alginates, powdered tragacanth, guar gum,cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate,carboxymethyl cellulose calcium, sodium carboxymethyl cellulose),polyvinyl pyrrolidone, methyl cellulose, pre-gelatinized starch,hydroxypropyl methyl cellulose, microcrystalline cellulose, and mixturesthereof.

Examples of suitable fillers for use in the pharmaceutical compositionsand dosage forms disclosed herein include, but are not limited to, talc,calcium carbonate (e.g., granules or powder), microcrystallinecellulose, powdered cellulose, dextrates, kaolin, mannitol, silicicacid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof.

Disintegrants may be used in the compositions of the invention toprovide tablets that disintegrate when exposed to an aqueousenvironment. Too much of a disintegrant may produce tablets whichdisintegrate in the bottle. Too little may be insufficient fordisintegration to occur, thus altering the rate and extent of release ofthe active ingredients from the dosage form. Thus, a sufficient amountof disintegrant that is neither too little nor too much to detrimentallyalter the release of the active ingredient(s) may be used to form thedosage forms of the compounds disclosed herein. The amount ofdisintegrant used may vary based upon the type of formulation and modeof administration, and may be readily discernible to those of ordinaryskill in the art. About 0.5 to about 15 weight percent of disintegrant,or about 1 to about 5 weight percent of disintegrant, may be used in thepharmaceutical composition. Disintegrants that can be used to formpharmaceutical compositions and dosage forms of the invention include,but are not limited to, agar-agar, alginic acid, calcium carbonate,microcrystalline cellulose, croscarmellose sodium, crospovidone,polacrilin potassium, sodium starch glycolate, potato or tapioca starch,other starches, pre-gelatinized starch, other starches, clays, otheraligns, other celluloses, gums or mixtures thereof.

Lubricants which can be used to form pharmaceutical compositions anddosage forms of the invention include, but are not limited to, calciumstearate, magnesium stearate, sodium stearyl fumarate, mineral oil,light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol,other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenatedvegetable oil (e.g., peanut oil, cottonseed oil, sunflower oil, sesameoil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate,ethyl aureate, agar, or mixtures thereof. Additional lubricants include,for example, a syloid silica gel, a coagulated aerosol of syntheticsilica, silicified microcrystalline cellulose, or mixtures thereof. Alubricant can optionally be added in an amount of less than about 0.5%or less than about 1% (by weight) of the pharmaceutical composition.

When aqueous suspensions and/or elixirs are desired for oraladministration, the active pharmaceutical ingredient(s) may be combinedwith various sweetening or flavoring agents, coloring matter or dyesand, if so desired, emulsifying and/or suspending agents, together withsuch diluents as water, ethanol, propylene glycol, glycerin and variouscombinations thereof.

The tablets can be uncoated or coated by known techniques to delaydisintegration and absorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period. For example, a timedelay material such as glyceryl monostearate or glyceryl distearate canbe employed. Formulations for oral use can also be presented as hardgelatin capsules wherein the active ingredient is mixed with an inertsolid diluent, for example, calcium carbonate, calcium phosphate orkaolin, or as soft gelatin capsules wherein the active ingredient ismixed with water or an oil medium, for example, peanut oil, liquidparaffin or olive oil.

Surfactants which can be used to form pharmaceutical compositions anddosage forms of the invention include, but are not limited to,hydrophilic surfactants, lipophilic surfactants, and mixtures thereof.That is, a mixture of hydrophilic surfactants may be employed, a mixtureof lipophilic surfactants may be employed, or a mixture of at least onehydrophilic surfactant and at least one lipophilic surfactant may beemployed.

A suitable hydrophilic surfactant may generally have an HLB value of atleast 10, while suitable lipophilic surfactants may generally have anHLB value of or less than about 10. An empirical parameter used tocharacterize the relative hydrophilicity and hydrophobicity of non-ionicamphiphilic compounds is the hydrophilic-lipophilic balance (“HLB”value). Surfactants with lower HLB values are more lipophilic orhydrophobic, and have greater solubility in oils, while surfactants withhigher HLB values are more hydrophilic, and have greater solubility inaqueous solutions. Hydrophilic surfactants are generally considered tobe those compounds having an HLB value greater than about 10, as well asanionic, cationic, or zwitterionic compounds for which the HLB scale isnot generally applicable. Similarly, lipophilic (i.e., hydrophobic)surfactants are compounds having an HLB value equal to or less thanabout 10. However, HLB value of a surfactant is merely a rough guidegenerally used to enable formulation of industrial, pharmaceutical andcosmetic emulsions.

Hydrophilic surfactants may be either ionic or non-ionic. Suitable ionicsurfactants include, but are not limited to, alkylammonium salts;fusidic acid salts; fatty acid derivatives of amino acids,oligopeptides, and polypeptides; glyceride derivatives of amino acids,oligopeptides, and polypeptides; lecithins and hydrogenated lecithins;lysolecithins and hydrogenated lysolecithins; phospholipids andderivatives thereof lysophospholipids and derivatives thereof; carnitinefatty acid ester salts; salts of alkylsulfates; fatty acid salts; sodiumdocusate; acylactylates; mono- and di-acetylated tartaric acid esters ofmono- and di-glycerides; succinylated mono- and di-glycerides; citricacid esters of mono- and di-glycerides; and mixtures thereof.

Within the aforementioned group, ionic surfactants include, by way ofexample: lecithins, lysolecithin, phospholipids, lysophospholipids andderivatives thereof; carnitine fatty acid ester salts; salts ofalkylsulfates; fatty acid salts; sodium docusate; acylactylates; mono-and di-acetylated tartaric acid esters of mono- and di-glycerides;succinylated mono- and di-glycerides; citric acid esters of mono- anddi-glycerides; and mixtures thereof.

Ionic surfactants may be the ionized forms of lecithin, lysolecithin,phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol,phosphatidic acid, phosphatidylserine, lysophosphatidylcholine,lysophosphatidylethanolamine, lysophosphatidylglycerol, lysophosphatidicacid, lysophosphatidylserine, PEG-phosphatidylethanolamine,PVP-phosphatidylethanolamine, lactylic esters of fatty acids,stearoyl-2-lactylate, stearoyl lactylate, succinylated monoglycerides,mono/diacetylated tartaric acid esters of mono/diglycerides, citric acidesters of mono/diglycerides, cholylsarcosine, caproate, caprylate,caprate, laurate, myristate, palmitate, oleate, ricinoleate, linoleate,linolenate, stearate, lauryl sulfate, teracecyl sulfate, docusate,lauroyl carnitines, palmitoyl carnitines, myristoyl carnitines, andsalts and mixtures thereof.

Hydrophilic non-ionic surfactants may include, but not limited to,alkylglucosides; alkylmaltosides; alkylthioglucosides; laurylmacrogolglycerides; polyoxyalkylene alkyl ethers such as polyethyleneglycol alkyl ethers; polyoxyalkylene alkylphenols such as polyethyleneglycol alkyl phenols; polyoxyalkylene alkyl phenol fatty acid esterssuch as polyethylene glycol fatty acids monoesters and polyethyleneglycol fatty acids diesters; polyethylene glycol glycerol fatty acidesters; polyglycerol fatty acid esters; polyoxyalkylene sorbitan fattyacid esters such as polyethylene glycol sorbitan fatty acid esters;hydrophilic transesterification products of a polyol with at least onemember of the group consisting of glycerides, vegetable oils,hydrogenated vegetable oils, fatty acids, and sterols; polyoxyethylenesterols, derivatives, and analogs thereof; polyoxyethylated vitamins andderivatives thereof; polyoxyethylene-polyoxypropylene block copolymers;and mixtures thereof; polyethylene glycol sorbitan fatty acid esters andhydrophilic transesterification products of a polyol with at least onemember of the group consisting of triglycerides, vegetable oils, andhydrogenated vegetable oils. The polyol may be glycerol, ethyleneglycol, polyethylene glycol, sorbitol, propylene glycol,pentaerythritol, or a saccharide.

Other hydrophilic-non-ionic surfactants include, without limitation,PEG-10 laurate, PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-32dilaurate, PEG-12 oleate, PEG-15 oleate, PEG-20 oleate, PEG-20 dioleate,PEG-32 oleate, PEG-200 oleate, PEG-400 oleate, PEG-15 stearate, PEG-32distearate, PEG-40 stearate, PEG-100 stearate, PEG-20 dilaurate, PEG-25glyceryl trioleate, PEG-32 dioleate, PEG-20 glyceryl laurate, PEG-30glyceryl laurate, PEG-20 glyceryl stearate, PEG-20 glyceryl oleate,PEG-30 glyceryl oleate, PEG-30 glyceryl laurate, PEG-40 glyceryllaurate, PEG-40 palm kernel oil, PEG-50 hydrogenated castor oil, PEG-40castor oil, PEG-35 castor oil, PEG-60 castor oil, PEG-40 hydrogenatedcastor oil, PEG-60 hydrogenated castor oil, PEG-60 corn oil, PEG-6caprate/caprylate glycerides, PEG-8 caprate/caprylate glycerides,polyglyceryl-10 laurate, PEG-30 cholesterol, PEG-25 phytosterol, PEG-30soya sterol, PEG-20 trioleate, PEG-40 sorbitan oleate, PEG-80 sorbitanlaurate, polysorbate 20, polysorbate 80, POE-9 lauryl ether, POE-23lauryl ether, POE-10 oleyl ether, POE-20 oleyl ether, POE-20 stearylether, tocopheryl PEG-100 succinate, PEG-24 cholesterol, polyglyceryl-10oleate, Tween 40, Tween 60, sucrose monostearate, sucrose monolaurate,sucrose monopalmitate, PEG 10-100 nonyl phenol series, PEG 15-100 octylphenol series, and poloxamers.

Suitable lipophilic surfactants include, by way of example only: fattyalcohols; glycerol fatty acid esters; acetylated glycerol fatty acidesters; lower alcohol fatty acids esters; propylene glycol fatty acidesters; sorbitan fatty acid esters; polyethylene glycol sorbitan fattyacid esters; sterols and sterol derivatives; polyoxyethylated sterolsand sterol derivatives; polyethylene glycol alkyl ethers; sugar esters;sugar ethers; lactic acid derivatives of mono- and di-glycerides;hydrophobic transesterification products of a polyol with at least onemember of the group consisting of glycerides, vegetable oils,hydrogenated vegetable oils, fatty acids and sterols; oil-solublevitamins/vitamin derivatives; and mixtures thereof. Within this group,preferred lipophilic surfactants include glycerol fatty acid esters,propylene glycol fatty acid esters, and mixtures thereof, or arehydrophobic transesterification products of a polyol with at least onemember of the group consisting of vegetable oils, hydrogenated vegetableoils, and triglycerides.

In an embodiment, the composition may include a solubilizer to ensuregood solubilization and/or dissolution of a compound described hereinand to minimize precipitation of the compound. This can be especiallyimportant for compositions for non-oral use—e.g., compositions forinjection. A solubilizer may also be added to increase the solubility ofthe hydrophilic drug and/or other components, such as surfactants, or tomaintain the composition as a stable or homogeneous solution ordispersion.

Examples of suitable solubilizers include, but are not limited to, thefollowing: alcohols and polyols, such as ethanol, isopropanol, butanol,benzyl alcohol, ethylene glycol, propylene glycol, butanediols andisomers thereof, glycerol, pentaerythritol, sorbitol, mannitol,transcutol, dimethyl isosorbide, polyethylene glycol, polypropyleneglycol, polyvinylalcohol, hydroxypropyl methylcellulose and othercellulose derivatives, cyclodextrins and cyclodextrin derivatives;ethers of polyethylene glycols having an average molecular weight ofabout 200 to about 6000, such as tetrahydrofurfuryl alcohol PEG ether(glycofurol) or methoxy PEG; amides and other nitrogen-containingcompounds such as 2-pyrrolidone, 2-piperidone, ε-caprolactam,N-alkylpyrrolidone, N-hydroxyalkylpyrrolidone, N-alkylpiperidone,N-alkylcaprolactam, dimethylacetamide and polyvinylpyrrolidone; esterssuch as ethyl propionate, tributylcitrate, acetyl triethylcitrate,acetyl tributyl citrate, triethylcitrate, ethyl oleate, ethyl caprylate,ethyl butyrate, triacetin, propylene glycol monoacetate, propyleneglycol diacetate, ε-caprolactone and isomers thereof, δ-valerolactoneand isomers thereof, β-butyrolactone and isomers thereof; and othersolubilizers known in the art, such as dimethyl acetamide, dimethylisosorbide, N-methyl pyrrolidones, monooctanoin, diethylene glycolmonoethyl ether, and water.

Mixtures of solubilizers may also be used. Examples include, but notlimited to, triacetin, triethylcitrate, ethyl oleate, ethyl caprylate,dimethylacetamide, N-methylpyrrolidone, N-hydroxyethylpyrrolidone,polyvinylpyrrolidone, hydroxypropyl methylcellulose, hydroxypropylcyclodextrins, ethanol, polyethylene glycol 200-100, glycofurol,transcutol, propylene glycol, and dimethyl isosorbide. Particularlypreferred solubilizers include sorbitol, glycerol, triacetin, ethylalcohol, PEG-400, glycofurol and propylene glycol.

The amount of solubilizer that can be included is not particularlylimited. The amount of a given solubilizer may be limited to abioacceptable amount, which may be readily determined by one of skill inthe art. In some circumstances, it may be advantageous to includeamounts of solubilizers far in excess of bioacceptable amounts, forexample to maximize the concentration of the drug, with excesssolubilizer removed prior to providing the composition to a patientusing conventional techniques, such as distillation or evaporation.Thus, if present, the solubilizer can be in a weight ratio of 10%, 25%,50%, 100%, or up to about 200% by weight, based on the combined weightof the drug, and other excipients. If desired, very small amounts ofsolubilizer may also be used, such as 5%, 2%, 1% or even less.Typically, the solubilizer may be present in an amount of about 1% toabout 100%, more typically about 5% to about 25% by weight.

The composition can further include one or more pharmaceuticallyacceptable additives and excipients. Such additives and excipientsinclude, without limitation, detackifiers, anti-foaming agents,buffering agents, polymers, antioxidants, preservatives, chelatingagents, viscomodulators, tonicifiers, flavorants, colorants, odorants,opacifiers, suspending agents, binders, fillers, plasticizers,lubricants, and mixtures thereof.

In addition, an acid or a base may be incorporated into the compositionto facilitate processing, to enhance stability, or for other reasons.Examples of pharmaceutically acceptable bases include amino acids, aminoacid esters, ammonium hydroxide, potassium hydroxide, sodium hydroxide,sodium hydrogen carbonate, aluminum hydroxide, calcium carbonate,magnesium hydroxide, magnesium aluminum silicate, synthetic aluminumsilicate, synthetic hydrocalcite, magnesium aluminum hydroxide,diisopropylethylamine, ethanolamine, ethylenediamine, triethanolamine,triethylamine, triisopropanolamine, trimethylamine,tris(hydroxymethyl)aminomethane (TRIS) and the like. Also suitable arebases that are salts of a pharmaceutically acceptable acid, such asacetic acid, acrylic acid, adipic acid, alginic acid, alkanesulfonicacid, amino acids, ascorbic acid, benzoic acid, boric acid, butyricacid, carbonic acid, citric acid, fatty acids, formic acid, fumaricacid, gluconic acid, hydroquinosulfonic acid, isoascorbic acid, lacticacid, maleic acid, oxalic acid, para-bromophenylsulfonic acid, propionicacid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinicacid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonicacid, uric acid, and the like. Salts of polyprotic acids, such as sodiumphosphate, disodium hydrogen phosphate, and sodium dihydrogen phosphatecan also be used. When the base is a salt, the cation can be anyconvenient and pharmaceutically acceptable cation, such as ammonium,alkali metals and alkaline earth metals. Example may include, but notlimited to, sodium, potassium, lithium, magnesium, calcium and ammonium.

Suitable acids are pharmaceutically acceptable organic or inorganicacids. Examples of suitable inorganic acids include hydrochloric acid,hydrobromic acid, hydriodic acid, sulfuric acid, nitric acid, boricacid, phosphoric acid, and the like. Examples of suitable organic acidsinclude acetic acid, acrylic acid, adipic acid, alginic acid,alkanesulfonic acids, amino acids, ascorbic acid, benzoic acid, boricacid, butyric acid, carbonic acid, citric acid, fatty acids, formicacid, fumaric acid, gluconic acid, hydroquinosulfonic acid, isoascorbicacid, lactic acid, maleic acid, methanesulfonic acid, oxalic acid,para-bromophenylsulfonic acid, propionic acid, p-toluenesulfonic acid,salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid,thioglycolic acid, toluenesulfonic acid and uric acid.

Pharmaceutical Compositions for Injection

In some embodiments, a pharmaceutical composition is provided forinjection containing an active pharmaceutical ingredient or combinationof active pharmaceutical ingredients, such as an HDAC inhibitor (e.g.,such as a selective HDAC inhibitor, including one or more compounds offormulas I, II, III, IV, V, VI, VII, and/or VIII), and a pharmaceuticalexcipient suitable for injection.

The forms in which the compositions described herein may be incorporatedfor administration by injection include aqueous or oil suspensions, oremulsions, with sesame oil, corn oil, cottonseed oil, or peanut oil, aswell as elixirs, mannitol, dextrose, or a sterile aqueous solution, andsimilar pharmaceutical vehicles.

Aqueous solutions in saline are also conventionally used for injection.Ethanol, glycerol, propylene glycol and liquid polyethylene glycol (andsuitable mixtures thereof), cyclodextrin derivatives, and vegetable oilsmay also be employed. The proper fluidity can be maintained, forexample, by the use of a coating, such as lecithin, for the maintenanceof the required particle size in the case of dispersion and by the useof surfactants. The prevention of the action of microorganisms can bebrought about by various antibacterial and antifungal agents, forexample, parabens, chlorobutanol, phenol, sorbic acid, and thimerosal.

Sterile injectable solutions are prepared by incorporating an activepharmaceutical ingredient or combination of active pharmaceuticalingredients in the required amounts in the appropriate solvent withvarious other ingredients as enumerated above, as required, followed byfiltered sterilization. Generally, dispersions are prepared byincorporating the various sterilized active ingredients into a sterilevehicle which contains the basic dispersion medium and the requiredother ingredients from those enumerated above. In the case of sterilepowders for the preparation of sterile injectable solutions, certaindesirable methods of preparation are vacuum-drying and freeze-dryingtechniques which yield a powder of the active ingredient plus anyadditional desired ingredient from a previously sterile-filteredsolution thereof.

Pharmaceutical Compositions for Topical Delivery

In some embodiments, a pharmaceutical composition is provided fortransdermal delivery containing an active pharmaceutical ingredient orcombination of active pharmaceutical ingredients, such as HDACinhibitors described herein (e.g., such as a selective HDAC inhibitor,including one or more compounds of formulas I, II, III, IV, V, VI, VII,and/or VIII), and a pharmaceutical excipient suitable for transdermaldelivery.

Compositions described herein can be formulated into preparations insolid, semi-solid, or liquid forms suitable for local or topicaladministration, such as gels, water soluble jellies, creams, lotions,suspensions, foams, powders, slurries, ointments, solutions, oils,pastes, suppositories, sprays, emulsions, saline solutions,dimethylsulfoxide (DMSO)-based solutions. In general, carriers withhigher densities are capable of providing an area with a prolongedexposure to the active ingredients. In contrast, a solution formulationmay provide more immediate exposure of the active ingredient to thechosen area.

The pharmaceutical compositions also may comprise suitable solid or gelphase carriers or excipients, which are compounds that allow increasedpenetration of, or assist in the delivery of, therapeutic moleculesacross the stratum corneum permeability barrier of the skin. There aremany of these penetration-enhancing molecules known to those trained inthe art of topical formulation. Examples of such carriers and excipientsinclude, but are not limited to, humectants (e.g., urea), glycols (e.g.,propylene glycol), alcohols (e.g., ethanol), fatty acids (e.g., oleicacid), surfactants (e.g., isopropyl myristate and sodium laurylsulfate), pyrrolidones, glycerol monolaurate, sulfoxides, terpenes(e.g., menthol), amines, amides, alkanes, alkanols, water, calciumcarbonate, calcium phosphate, various sugars, starches, cellulosederivatives, gelatin, and polymers such as polyethylene glycols.

Another exemplary formulation for use in the methods described hereinemploys transdermal delivery devices (“patches”). Such transdermalpatches may be used to provide continuous or discontinuous infusion ofan active pharmaceutical ingredient or combination of activepharmaceutical ingredients in controlled amounts, either with or withoutanother active pharmaceutical ingredient.

The construction and use of transdermal patches for the delivery ofpharmaceutical agents is well known in the art. See, e.g., U.S. Pat.Nos. 5,023,252; 4,992,445; and 5,001,139, the entirety of which areincorporated herein by reference. Such patches may be constructed forcontinuous, pulsatile, or on demand delivery of pharmaceutical agents.

Pharmaceutical Compositions for Inhalation

Compositions for inhalation or insufflation include solutions andsuspensions in pharmaceutically acceptable, aqueous or organic solvents,or mixtures thereof, and powders. The liquid or solid compositions maycontain suitable pharmaceutically acceptable excipients as describedsupra and the HDAC inhibitors described herein (e.g., such as aselective HDAC inhibitor, including one or more compounds of formulas I,II, III, IV, V, VI, VII, and/or VIII). Preferably the compositions areadministered by the oral or nasal respiratory route for local orsystemic effect. Compositions in preferably pharmaceutically acceptablesolvents may be nebulized by use of inert gases. Nebulized solutions maybe inhaled directly from the nebulizing device or the nebulizing devicemay be attached to a face mask tent, or intermittent positive pressurebreathing machine. Solution, suspension, or powder compositions may beadministered, preferably orally or nasally, from devices that deliverthe formulation in an appropriate manner. Dry powder inhalers may alsobe used to provide inhaled delivery of the compositions.

Other Pharmaceutical Compositions

Pharmaceutical compositions of the HDAC inhibitors described herein mayalso be prepared from compositions described herein and one or morepharmaceutically acceptable excipients suitable for sublingual, buccal,rectal, intraosseous, intraocular, intranasal, epidural, or intraspinaladministration. Preparations for such pharmaceutical compositions arewell-known in the art. See, e.g., Anderson, Philip O.; Knoben, James E.;Troutman, William G, eds., Handbook of Clinical Drug Data, TenthEdition, McGraw-Hill, 2002; and Pratt and Taylor, eds., Principles ofDrug Action, Third Edition, Churchill Livingston, N.Y., 1990, each ofwhich is incorporated by reference herein in its entirety.

Administration of an active pharmaceutical ingredient or combination ofactive pharmaceutical ingredients or a pharmaceutical compositionthereof can be effected by any method that enables delivery of thecompounds to the site of action. These methods include oral routes,intraduodenal routes, parenteral injection (including intravenous,intraarterial, subcutaneous, intramuscular, intravascular,intraperitoneal or infusion), topical (e.g., transdermal application),rectal administration, via local delivery by catheter or stent orthrough inhalation. The active pharmaceutical ingredient or combinationof active pharmaceutical ingredients can also be administeredintraadiposally or intrathecally.

Exemplary parenteral administration forms include solutions orsuspensions of active compound in sterile aqueous solutions, forexample, aqueous propylene glycol or dextrose solutions. Such dosageforms can be suitably buffered, if desired.

The invention also provides kits. The kits include an activepharmaceutical ingredient or combination of active pharmaceuticalingredients, either alone or in combination in suitable packaging, andwritten material that can include instructions for use, discussion ofclinical studies and listing of side effects. Such kits may also includeinformation, such as scientific literature references, package insertmaterials, clinical trial results, and/or summaries of these and thelike, which indicate or establish the activities and/or advantages ofthe composition, and/or which describe dosing, administration, sideeffects, drug interactions, or other information useful to the healthcare provider. Such information may be based on the results of variousstudies, for example, studies using experimental animals involving invivo models and studies based on human clinical trials. The kit mayfurther contain another active pharmaceutical ingredient. In selectedembodiments, an active pharmaceutical ingredient or combination ofactive pharmaceutical ingredients are provided as separate compositionsin separate containers within the kit. In selected embodiments, anactive pharmaceutical ingredient or combination of active pharmaceuticalingredients are provided as a single composition within a container inthe kit. Suitable packaging and additional articles for use (e.g.,measuring cup for liquid preparations, foil wrapping to minimizeexposure to air, and the like) are known in the art and may be includedin the kit. Kits described herein can be provided, marketed and/orpromoted to health providers, including physicians, nurses, pharmacists,formulary officials, and the like. Kits may also, in selectedembodiments, be marketed directly to the consumer.

In some embodiments, the invention provides a kit comprising acomposition comprising a therapeutically effective amount of an activepharmaceutical ingredient (e.g., HDAC inhibitor) or combination ofactive pharmaceutical ingredients or a pharmaceutically acceptable salt,solvate, hydrate, cocrystal, or prodrug thereof. These compositions aretypically pharmaceutical compositions. The kit is for co-administrationof the active pharmaceutical ingredient or combination of activepharmaceutical ingredients, either simultaneously or separately.

In some embodiments, the invention provides a kit comprising (1) acomposition comprising a therapeutically effective amount of an activepharmaceutical ingredient (e.g., HDAC inhibitor) or combination ofactive pharmaceutical ingredients or a pharmaceutically acceptable salt,solvate, hydrate, cocrystal, or prodrug thereof, and (2) a diagnostictest for determining whether a patient's cancer is a particular subtypeof a cancer. Any of the foregoing diagnostic methods may be utilized inthe kit.

For example, a kit of the invention may include one or more HDACinhibitors and a chemotherapeutic agent and/or an immunotherapeuticagent.

The kits described above are preferably for use in the treatment of thediseases and conditions described herein. In a particular embodiment,the kits are for use in the treatment of cancer, neurological disorders,or immunological disorders.

Dosages and Dosing Regimens

The amounts of the pharmaceutical compositions administered using themethods herein, such as the dosages of HDAC inhibitors, will bedependent on the human or mammal being treated, the severity of thedisorder or condition, the rate of administration, the disposition ofthe active pharmaceutical ingredients and the discretion of theprescribing physician. However, an effective dosage is in the range ofabout 0.001 to about 100 mg per kg body weight per day, such as about 1to about 35 mg/kg/day, in single or divided doses. For a 70 kg human,this would amount to about 0.05 to 7 g/day, such as about 0.05 to about2.5 g/day. In some instances, dosage levels below the lower limit of theaforesaid range may be more than adequate, while in other cases stilllarger doses may be employed without causing any harmful sideeffect—e.g., by dividing such larger doses into several small doses foradministration throughout the day. The dosage of the pharmaceuticalcompositions and active pharmaceutical ingredients may be provided inunits of mg/kg of body mass or in mg/m² of body surface area.

In some embodiments, a pharmaceutical composition or activepharmaceutical ingredient is administered in a single dose. Suchadministration may be by injection, e.g., intravenous injection, inorder to introduce the active pharmaceutical ingredient quickly.However, other routes, including the preferred oral route, may be usedas appropriate. A single dose of a pharmaceutical composition may alsobe used for treatment of an acute condition.

In some embodiments, a pharmaceutical composition or activepharmaceutical ingredient is administered in multiple doses. In anembodiment, a pharmaceutical composition is administered in multipledoses. Dosing may be once, twice, three times, four times, five times,six times, or more than six times per day. Dosing may be once a month,once every two weeks, once a week, or once every other day. In otherembodiments, a pharmaceutical composition is administered about once perday to about 6 times per day. In some embodiments, a pharmaceuticalcomposition is administered once daily, while in other embodiments, apharmaceutical composition is administered twice daily, and in otherembodiments a pharmaceutical composition is administered three timesdaily.

Administration of the active pharmaceutical ingredients may continue aslong as necessary. In selected embodiments, a pharmaceutical compositionis administered for more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 day(s). Insome embodiments, a pharmaceutical composition is administered for lessthan 28, 14, 7, 6, 5, 4, 3, 2, or 1 day(s). In some embodiments, apharmaceutical composition is administered chronically on an ongoingbasis—e.g., for the treatment of chronic effects. In some embodiments,the administration of a pharmaceutical composition continues for lessthan about 7 days. In yet another embodiment the administrationcontinues for more than about 6, 10, 14, 28 days, two months, sixmonths, or one year. In some cases, continuous dosing is achieved andmaintained as long as necessary.

In some embodiments, an effective dosage of an active pharmaceuticalingredient disclosed herein is in the range of about 1 mg to about 500mg, about 10 mg to about 300 mg, about 20 mg to about 250 mg, about 25mg to about 200 mg, about 10 mg to about 200 mg, about 20 mg to about150 mg, about 30 mg to about 120 mg, about 10 mg to about 90 mg, about20 mg to about 80 mg, about 30 mg to about 70 mg, about 40 mg to about60 mg, about 45 mg to about 55 mg, about 48 mg to about 52 mg, about 50mg to about 150 mg, about 60 mg to about 140 mg, about 70 mg to about130 mg, about 80 mg to about 120 mg, about 90 mg to about 110 mg, about95 mg to about 105 mg, about 150 mg to about 250 mg, about 160 mg toabout 240 mg, about 170 mg to about 230 mg, about 180 mg to about 220mg, about 190 mg to about 210 mg, about 195 mg to about 205 mg, or about198 to about 202 mg. In some embodiments, an effective dosage of anactive pharmaceutical ingredient disclosed herein is less than about 25mg, less than about 50 mg, less than about 75 mg, less than about 100mg, less than about 125 mg, less than about 150 mg, less than about 175mg, less than about 200 mg, less than about 225 mg, or less than about250 mg. In some embodiments, an effective dosage of an activepharmaceutical ingredient disclosed herein is greater than about 25 mg,greater than about 50 mg, greater than about 75 mg, greater than about100 mg, greater than about 125 mg, greater than about 150 mg, greaterthan about 175 mg, greater than about 200 mg, greater than about 225 mg,or greater than about 250 mg.

In some embodiments, an effective dosage of an active pharmaceuticalingredient disclosed herein is in the range of about 0.01 mg/kg to about200 mg/kg, or about 0.1 to 100 mg/kg, or about 1 to 50 mg/kg.

In some embodiments, an active pharmaceutical ingredient is adminsteredat a dosage of 10 to 200 mg BID, including 50, 60, 70, 80, 90, 100, 150,or 200 mg BID. In some embodiments, an active pharmaceutical ingredientis adminstered at a dosage of 10 to 500 mg BID, including 1, 5, 10, 15,25, 50, 75, 100, 150, 200, 300, 400, or 500 mg BID.

In some instances, dosage levels below the lower limit of the aforesaidranges may be more than adequate, while in other cases still largerdoses may be employed without causing any harmful side effect—e.g., bydividing such larger doses into several small doses for administrationthroughout the day. Of course, as those skilled in the art willappreciate, the dosage actually administered will depend upon thecondition being treated, the age, health and weight of the recipient,the type of concurrent treatment, if any, and the frequency oftreatment. Moreover, the effective dosage amount may be determined byone skilled in the art on the basis of routine empirical activitytesting to measure the bioactivity of the compound(s) in a bioassay, andthus establish the appropriate dosage to be administered.

An effective amount of the combination of the active pharmaceuticalingredient may be administered in either single or multiple doses by anyof the accepted modes of administration of agents having similarutilities, including rectal, buccal, intranasal and transdermal routes,by intra-arterial injection, intravenously, intraperitoneally,parenterally, intramuscularly, subcutaneously, orally, topically, or asan inhalant.

In some embodiments, the compositions described herein further includecontrolled-release, sustained release, or extended-release therapeuticdosage forms for administration of the compounds described herein, whichinvolves incorporation of the compounds into a suitable delivery systemin the formation of certain compositions. This dosage form controlsrelease of the compound(s) in such a manner that an effectiveconcentration of the compound(s) in the bloodstream may be maintainedover an extended period of time, with the concentration in the bloodremaining relatively constant, to improve therapeutic results and/orminimize side effects. Additionally, a controlled-release system wouldprovide minimum peak to trough fluctuations in blood plasma levels ofthe compound.

The following examples describe the invention in further detail. Theseexamples are provided for illustrative purposes only, and should in noway be considered as limiting the invention.

EXAMPLES Example 1: Compounds SP-2-213, SP-2-223, and SP-2-225 wereTested as Selective Inhibitors of HDAC6 as Compared to Pan-HDAC, HDAC1,and HDAC3

SP-2-213, SP-2-223, and SP-2-225 were tested in assay to determine theselectivity of the compounds for specific HDAC proteins. The compoundswere tested against Pan HDAC, HDAC1, HDAC3, and HDAC6 proteins in vitro.As shown in FIGS. 1 and 2A-2C, SP-2-213, SP-2-223, and SP-2-225 wereinhibitors of HDAC6.

However, SP-2-213, SP-2-223, and SP-2-225 were found to be selectiveinhibitors of HDAC6 as compared to Pan-HDACs, HDAC1, and HDAC3.

As compared to HDAC1 and HDAC3, SP-2-213 was about 4 times and about 12times more selective for HDAC6, respectively.

As compared to HDAC1 and HDAC3, SP-2-223 was about 44 times and about 18times more selective for HDAC6, respectively.

As compared to HDAC1 and HDAC3, SP-2-225 was about 65 times and about 22times more selective for HDAC6, respectively.

A number of patent and non-patent publications are cited herein in orderto describe the state of the art to which this invention pertains. Theentire disclosure of each of these publications is incorporated byreference herein.

While certain embodiments have been described and/or exemplified above,various other embodiments will be apparent to those skilled in the artfrom the foregoing disclosure. The invention described herein is,therefore, not limited to the particular embodiments described and/orexemplified, but is capable of considerable variation and modificationwithout departure from the scope of the appended claims.

Moreover, as used herein, the term “about” means that dimensions, sizes,formulations, parameters, shapes and other quantities andcharacteristics are not and need not be exact, but may be approximateand/or larger or smaller, as desired, reflecting tolerances, conversionfactors, rounding off, measurement error and the like, and other factorsknown to those of skill in the art. In general, a dimension, size,formulation, parameter, shape or other quantity or characteristic is“about” or “approximate” whether or not expressly stated to be such. Itis noted that embodiments of very different sizes, shapes and dimensionsmay employ the described arrangements.

Furthermore, the transitional terms “comprising”, “consistingessentially of” and “consisting of”, when used in the appended claims,in original and amended form, define the claim scope with respect towhat unrecited additional claim elements or steps, if any, are excludedfrom the scope of the claim(s). The term “comprising” is intended to beinclusive or open-ended and does not exclude any additional, unrecitedelement, method, step or material. The term “consisting of” excludes anyelement, step or material other than those specified in the claim and,in the latter instance, impurities ordinary associated with thespecified material(s). The term “consisting essentially of” limits thescope of a claim to the specified elements, steps or material(s) andthose that do not materially affect the basic and novelcharacteristic(s) of the claimed invention. All compounds, compositions,and methods described herein that embody the invention can, in alternateembodiments, be more specifically defined by any of the transitionalterms “comprising,” “consisting essentially of,” and “consisting of.”

What is claimed is:
 1. A compound of formula I, II, III, IV, or V:

wherein R¹ is H or optionally substituted alkyl or aryl; R² is H or optionally substituted alkyl or aryl; Q is a moiety selected from the group consisting of NR³—(CH₂)_(n)—, —(CH₂)_(n)—NR³—, NR³—C(═O)—(CH₂)_(n)—, —(CH₂)_(n)—C(═O)—NR³—, —C(═O)—NR³—(CH₂)_(n)—, and —(CH₂)_(n)—NR³—C(═O)—, wherein n is 0 or 1, and R³ is H or optionally substituted alkyl or aryl; each of R⁴, R⁵, R⁶, and R⁷ are independently selected from the group consisting of H, halo, and optionally substituted alkyl, aryl, alkoxy, and aryloxy; Z is O, S, S(═O), or S(═O)₂; A is a moiety selected from the group consisting of

or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, or prodrug thereof.
 2. The compound of claim 1, wherein the compound is of formula (VI):

or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, or prodrug thereof.
 3. The compound of claim 1, wherein the compound is of formula (VII):

or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, or prodrug thereof.
 4. The compound of claim 1, wherein the compound is of formula (VIII):

or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, or prodrug thereof.
 5. A pharmaceutical composition comprising a histone deacetylase (HDAC) inhibitor of claim 1, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, or prodrug thereof. 